Antagonists of lysophosphatidic acid receptors

ABSTRACT

Described herein are compounds that are antagonists of lysophosphatidic receptor(s). Also described are pharmaceutical compositions and medicaments that include the compounds described herein, as well as methods of using such antagonists, alone and in combination with other compounds, for treating LPA-dependent or LPA-mediated conditions or diseases.

RELATED APPLICATIONS

This application is a continuation application of U.S. Pat. No.8,048,902 entitled, “ANTAGONISTS OF LYSOPHOSPHATIDIC ACID RECEPTORS”filed on Dec. 15, 2008 now U.S. Pat. No. 8,048,902, which claims thebenefit of U.S. Provisional Application No. 61/122,568, entitled“ANTAGONISTS OF LYSOPHOSPHATIDIC ACID RECEPTORS” filed on Dec. 15, 2008,all of which are incorporated herein by reference in their entirety.

FIELD OF THE INVENTION

Described herein are compounds, methods of making such compounds,pharmaceutical compositions and medicaments comprising such compounds,and methods of using such compounds to treat, prevent or diagnosediseases, disorders or conditions associated with one or more of thelysophosphatidic acid (LPA) receptors.

BACKGROUND OF THE INVENTION

Lysophospholipids are membrane-derived bioactive lipid mediators.Lysophospholipids affect fundamental cellular functions that includeproliferation, differentiation, survival, migration, adhesion, invasion,and morphogensis. These functions influence many biological processesthat include, but are not limited to, neurogensis, angiogenesis, woundhealing, fibrosis, immunity, and carcinogenesis.

Lysophosphatidic acid (LPA) is a lysophospholipid that has been shown toact through sets of specific G protein-coupled receptors (GPCRs) in anautocrine and paracrine fashion. LPA binding to its cognate GPCRs (LPA₁,LPA₂, LPA₃, LPA₄, LPA₅, LPA₆) activates intracellular signaling pathwaysto produce a variety of biological responses. Antagonists of the LPAreceptors find use in the treatment of diseases, disorders or conditionsin which LPA plays a role.

SUMMARY OF THE INVENTION

In one aspect, presented herein are compounds of Formula (I) thatinhibit the physiological activity of lysophosphatidic acid (LPA), andtherefore, are useful as agents for the treatment or prevention ofdiseases in which inhibition of the physiological activity of LPA isuseful, such as diseases in which an LPA receptor participates, isinvolved in the etiology or pathology of the disease, or is otherwiseassociated with at least one symptom of the disease. In a relatedaspect, such compounds are useful as agents for the treatment orprevention of side effects, complications, or adverse events associatedwith the use of a different therapeutic agent or therapeutic action(e.g., radiation, surgery, etc) used in treating a disease or condition.

In one aspect, the compounds of Formula (I) are useful for the treatmentof fibrosis of organs (liver, kidney, lung, heart and the like), liverdiseases (acute hepatatis, chronic hepatitis, liver fibrosis, livercirrhosis, portal hypertension, regenerative failure, non-alcoholicsteatohepatitis (NASH), liver hypofunction, hepatic blood flow disorder,and the like), cell proliferative disease (cancer (solid tumor, solidtumor metastasis, vascular fibroma, myeloma, multiple myeloma, Kaposi'ssarcoma, leukemia, chronic lymphocytic leukemia (CLL) and the like) andinvasive metastasis of cancer cell, and the like), inflammatory disease(psoriasis, nephropathy, pneumonia and the like), gastrointestinal tractdisease (irritable bowel syndrome (IBS), inflammatory bowel disease(IBD), abnormal pancreatic secretion, and the like), renal disease,urinary tract-associated disease (benign prostatic hyperplasia orsymptoms associated with neuropathic bladder disease, spinal cord tumor,hernia of intervertebral disk, spinal canal stenosis, symptoms derivedfrom diabetes, lower urinary tract disease (obstruction of lower urinarytract, and the like), inflammatory disease of lower urinary tract,dysuria, frequent urination, and the like), pancreas disease, abnormalangiogenesis-associated disease (arterial obstruction and the like),scleroderma, brain-associated disease (cerebral infarction, cerebralhemorrhage, and the like), neuropathic pain, peripheral neuropathy, andthe like, ocular disease (age-related macular degeneration (AMD),diabetic retinopathy, proliferative vitreoretinopathy (PVR), cicatricialpemphigoid, glaucoma filtration surgery scarring, and the like). In oneaspect, the compounds of Formula (I) are used in the treatment offibrotic diseases or conditions.

In one aspect, described herein are compounds of Formula (I),pharmaceutically acceptable salts, solvates, and prodrugs thereof.Compounds of Formula (I) are antagonists of at least one of the LPAreceptors selected from LPA₁, LPA₂, LPA₃, LPA₄, LPA₅ and LPA₆. In oneembodiment, compounds of Formula (I) are antagonists of LPA₁. In oneembodiment, compounds of Formula (I) are antagonists of LPA₁ and/orLPA₃. In some embodiments, compounds of Formula (I) are antagonists ofLPA₁ and/or LPA₂. In some embodiments, compounds of Formula (I) areselective antagonists for one of the LPA receptors relative to the otherLPA receptors. In some embodiments, such a selective antagonist isselective for the LPA₁ receptor. In some embodiments, such a selectiveantagonist is selective for the LPA₂ receptor. In some embodiments, sucha selective antagonist is selective for the LPA₃ receptor.

Compounds of Formula (I) are used in the treatment of diseases,disorders, or conditions in which activation of at least one LPAreceptor by LPA contributes to the symptomology or progression of thedisease, disorder or condition. In one aspect, the methods, compounds,pharmaceutical compositions, and medicaments described herein compriseantagonists of LPA receptors. In one aspect, the methods, compounds,pharmaceutical compositions, and medicaments described herein compriseantagonists of LPA₁, LPA₂, or LPA₃, or combinations thereof.

In one aspect, provided herein is a compound of Formula (I), or apharmaceutically acceptable salt thereof:

wherein

-   -   R¹ is —CO₂H, —CO₂R^(D), —CN, tetrazolyl, —C(═O)NH₂, —C(═O)NHR¹⁰,        —C(═O)NHSO₂R¹⁰ or —C(═O)NHCH₂CH₂SO₃H; R^(D) is H or C₁-C₄alkyl;    -   L¹ is absent or C₁-C₆alkylene;    -   R³ is H, C₁-C₄alkyl, C₃-C₆cycloalkyl, or C₁-C₄-fluoroalkyl;    -   R⁷ is H or C₁-C₄alkyl;    -   R⁸ is H, C₁-C₄alkyl, or C₁-C₄-fluoroalkyl;    -   R¹⁰ is a C₁-C₆alkyl, C₁-C₆-fluoroalkyl, C₃-C₆cycloalkyl, or a        substituted or unsubstituted phenyl;    -   each of R^(A), R^(B), and R^(C) are independently selected from        H, F, Cl, Br, I, —CN, —OH, C₁-C₄alkyl, C₁-C₄-fluoroalkyl,        C₁-C₄-fluoroalkoxy, C₁-C₄alkoxy, and C₁-C₄heteroalkyl;    -   m is 0, 1, or 2; n is 0, 1, or 2; p is 0, 1, or 2.

In one aspect, provided is are compounds presented in Table 1, Table 2,Table 3, FIG. 1, FIG. 2, FIG. 3, FIG. 4, FIG. 5, FIG. 6, and FIG. 7.

Compounds of Formula (I) are antagonists of at least one LPA receptor.In some embodiments, the compound of Formula (I) is an antagonist ofLPA₁. In some embodiments, the compound of Formula (I) is an antagonistof LPA₂. In some embodiments, the compound of Formula (I) is anantagonist of LPA₃.

In some embodiments, presented herein are compounds selected from activemetabolites, tautomers, solvates, pharmaceutically acceptable salts orprodrugs of a compound of Formula (I).

In some embodiments, provided is a pharmaceutical composition comprisinga therapeutically effective amount of a compound of Formula (I). In someembodiments, the pharmaceutical composition also contains at least onepharmaceutically acceptable inactive ingredient.

In some embodiments, provided is a pharmaceutical composition comprisinga therapeutically effective amount of a compound of Formula (I), or apharmaceutically acceptable salt thereof, and at least onepharmaceutically acceptable inactive ingredient. In one aspect, thepharmaceutical composition is formulated for intravenous injection,subcutaneous injection, oral administration, inhalation, nasaladministration, topical administration, ophthalmic administration orotic administration. In some embodiments, the pharmaceutical compositionis a tablet, a pill, a capsule, a liquid, an inhalant, a nasal spraysolution, a suppository, a suspension, a gel, a colloid, a dispersion, asuspension, a solution, an emulsion, an ointment, a lotion, an eye dropor an ear drop.

In some embodiments, the pharmaceutical composition further comprisesone or more additional therapeutically active agents selected from:corticosteroids, immunosuppresants, analgesics, anti-cancer agent,anti-inflammatories, chemokine receptor antagonists, bronchodilators,leukotriene receptor antagonists, leukotriene formation inhibitors,monoacylglycerol kinase inhibitors, phospholipase A₁ inhibitors,phospholipase A₂ inhibitors, and lysophospholipase D (lysoPLD)inhibitors, autotaxin inhibitors, decongestants, antihistamines,mucolytics, anticholinergics, antitussives, expectorants, and β-2agonists.

In some embodiments, provided is a method comprising administering acompound of Formula (I) to a human with a LPA-dependent or LPA-mediateddisease or condition. In some embodiments, the human is already beingadministered one or more additional therapeutically active agents otherthan a compound of Formula (I). In some embodiments, the method furthercomprises administering one or more additional therapeutically activeagents other than a compound of Formula (I).

In some embodiments, the one or more additional therapeutically activeagents other than a compound of Formula (I) are selected from:corticosteroids, immunosuppresants, analgesics, anti-cancer agent,anti-inflammatories, chemokine receptor antagonists, bronchodilators,leukotriene receptor antagonists, leukotriene formation inhibitors,monoacylglycerol kinase inhibitors, phospholipase A₁ inhibitors,phospholipase A₂ inhibitors, and lysophospholipase D (lysoPLD)inhibitors, autotaxin inhibitors, decongestants, antihistamines,mucolytics, anticholinergics, antitussives, expectorants, and β-2agonists.

In another aspect is the use of a compound of Formula (I) in themanufacture of a medicament for treating a disease, disorder orcondition in which the activity of at least one LPA receptor contributesto the pathology and/or symptoms of the disease or condition. In oneembodiment of this aspect, the LPA receptor is selected from LPA₁, LPA₂,LPA₃, LPA₄, LPA₅ and LPA₆. In some embodiments, the LPA receptor isLPA₁. In some embodiments, the LPA receptor is LPA₂. In someembodiments, the LPA receptor is LPA₃. In some embodiments, the diseaseor condition is any of the diseases or conditions specified herein.

Also provided is a method of inhibiting the physiological activity ofLPA in a mammal comprising administering a therapeutically effectiveamount of a compound of Formula (I) or a pharmaceutically acceptablesalt thereof to the mammal in need thereof.

In one aspect, provided is a medicament for treating a LPA-dependent orLPA-mediated disease or condition in a mammal comprising atherapeutically effective amount of a compound of Formula (I).

In some cases disclosed herein is the use of a compound of Formula (I)in the manufacture of a medicament for the treatment of a LPA-dependentor LPA-mediated disease or condition.

In some cases disclosed herein is the use of a compound of Formula (I)in the treatment or prevention of a LPA-dependent or LPA-mediateddisease or condition.

In one aspect, is a method for treating or preventing a LPA-dependent orLPA-mediated disease or condition in a mammal comprising administering atherapeutically effective amount of a compound of Formula (I).

In one aspect, LPA-dependent or LPA-mediated diseases or conditionsinclude, but are not limited to, fibrosis of organs or tissues,scarring, liver diseases, dermatological conditions, cancer,cardiovascular disease, respiratory diseases or conditions, inflammatorydisease, gastrointestinal tract disease, renal disease, urinarytract-associated disease, inflammatory disease of lower urinary tract,dysuria, frequent urination, pancreas disease, arterial obstruction,cerebral infarction, cerebral hemorrhage, pain, peripheral neuropathy,and fibromyalgia.

In some embodiments, the LPA-dependent or LPA-mediated disease orcondition is selected from idiopathic pulmonary fibrosis; other diffuseparenchymal lung diseases of different etiologies including iatrogenicdrug-induced fibrosis, occupational and/or environmental inducedfibrosis, granulomatous diseases (sarcoidosis, hypersensitivitypneumonia), collagen vascular disease, alveolar proteinosis, langerhanscell granulomatosis, lymphangioleiomyomatosis, inherited diseases(Hermansky-Pudlak Syndrome, tuberous sclerosis, neurofibromatosis,metabolic storage disorders, familial interstitial lung disease);radiation induced fibrosis; chronic obstructive pulmonary disease(COPD); scleroderma; bleomycin induced pulmonary fibrosis; chronicasthma; silicosis; asbestos induced pulmonary fibrosis; acuterespiratory distress syndrome (ARDS); kidney fibrosis;tubulointerstitium fibrosis; glomerular nephritis; focal segmentalglomerular sclerosis; IgA nephropathy; hypertension; Alport; gutfibrosis; liver fibrosis; cirrhosis; alcohol induced liver fibrosis;toxic/drug induced liver fibrosis; hemochromatosis; nonalcoholicsteatohepatitis (NASH); biliary duct injury; primary biliary cirrhosis;infection induced liver fibrosis; viral induced liver fibrosis; andautoimmune hepatitis; corneal scarring; hypertrophic scarring; Duputrendisease, keloids, cutaneous fibrosis; cutaneous scleroderma; spinal cordinjury/fibrosis; myelofibrosis; vascular restenosis; atherosclerosis;arteriosclerosis; Wegener's granulomatosis; Peyronie's disease, chroniclymphocytic leukemia, tumor metastasis, transplant organ rejection,endometreosis, neonatal respiratory distress syndrome and neuropathicpain.

In one aspect, provided is a method for the treatment or prevention oforgan fibrosis in a mammal comprising administering a therapeuticallyeffective amount of a compound of Formula (I) or a pharmaceuticallyacceptable salt thereof to a mammal in need thereof.

In some embodiments, the organ fibrosis comprises lung fibrosis, renalfibrosis, or hepatic fibrosis.

In one aspect, provided is a method of improving lung function in amammal comprising administering a therapeutically effective amount of acompound of Formula (I) or a pharmaceutically acceptable salt thereof tothe mammal in need thereof. In one aspect, the mammal has been diagnosedas having lung fibrosis.

In one aspect, compounds disclosed herein are used to treat idiopathicpulmonary fibrosis (usual interstitial pneumonia) in a mammal.

In some embodiments, compounds disclosed herein are used to treatdiffuse parenchymal interstitial lung diseases in mammal: iatrogenicdrug induced, occupational/environmental (Farmer lung), granulomatousdiseases (sarcoidosis, hypersensitivity pneumonia), collagen vasculardisease (scleroderma and others), alveolar proteinosis, langerhans cellgranulonmatosis, lymphangioleiomyomatosis, Hermansky-Pudlak Syndrome,Tuberous sclerosis, neurofibromatosis, metabolic storage disorders,familial interstitial lung disease.

In some embodiments, compounds disclosed herein are used to treatpost-transplant fibrosis associated with chronic rejection in a mammal.Bronchiolitis obliterans for lung transplant.

In some embodiments, compounds disclosed herein are used to treatcutaneous fibrosis in a mammal: cutaneous scleroderma, Dupuytrendisease, keloids.

In one aspect, compounds disclosed herein are used to treat hepaticfibrosis with or without cirrhosis in a mammal: toxic/drug induced(hemochromatosis), alcoholic liver disease, viral hepatitis (hepatitis Bvirus, hepatitis C virus, HCV), nonalcoholic liver disease (NASH),metabolic and auto-immune.

In one aspect, compounds disclosed herein are used to treat renalfibrosis in a mammal: tubulointerstitium fibrosis, glomerular sclerosis.

In any of the aforementioned aspects involving the treatment of LPAdependent diseases or conditions are further embodiments comprisingadministering at least one additional agent in addition to theadministration of a compound having the structure of Formula (I). Invarious embodiments, each agent is administered in any order, includingsimultaneously.

In any of the embodiments disclosed herein, the mammal is a human.

In some embodiments, compounds provided herein are administered to ahuman. In some embodiments, compounds provided herein are orallyadministered to a human.

In some embodiments, compounds provided herein are used as antagonistsof at least one LPA receptor. In some embodiments, compounds providedherein are used for inhibiting the activity of at least one LPA receptoror for the treatment of a disease or condition that would benefit frominhibition of the activity of at least one LPA receptor. In one aspect,the LPA receptor is LPA₁.

In other embodiments, compounds provided herein are used for theformulation of a medicament for the inhibition of LPA₁ activity.

Other objects, features and advantages of the compounds, methods andcompositions described herein will become apparent from the followingdetailed description. It should be understood, however, that thedetailed description and the specific examples, while indicatingspecific embodiments, are given by way of illustration only, sincevarious changes and modifications within the spirit and scope of theinstant disclosure will become apparent to those skilled in the art fromthis detailed description.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1. Illustrative examples of compounds described herein.

FIG. 2. Illustrative examples of compounds described herein.

FIG. 3. Illustrative examples of compounds described herein.

FIG. 4. Illustrative examples of compounds described herein.

FIG. 5. Illustrative examples of compounds described herein.

FIG. 6. Illustrative examples of compounds described herein.

FIG. 7. Illustrative examples of compounds described herein.

DETAILED DESCRIPTION OF THE INVENTION

Lysophospholipids (such as lysophosphatidic acid (LPA)) affectfundamental cellular functions that include cellular proliferation,differentiation, survival, migration, adhesion, invasion, andmorphogensis. These functions influence many biological processes thatinclude neurogensis, angiogenesis, wound healing, immunity, andcarcinogenesis.

LPA acts through sets of specific G protein-coupled receptors (GPCRs) inan autocrine and paracrine fashion. LPA binding to its cognate GPCRs(LPA₁, LPA₂, LPA₃, LPA₄, LPA₅, LPA₆) activates intracellular signalingpathways to produce a variety of biological responses.

LPA has a role as a biological effector molecule, and has a diverserange of physiological actions such as, but not limited to, effects onblood pressure, platelet activation, and smooth muscle contraction, anda variety of cellular effects, which include cell growth, cell rounding,neurite retraction, and actin stress fiber formation and cell migration.The effects of LPA are predominantly receptor mediated.

Activation of the LPA receptors (LPA₁, LPA₂, LPA₃, LPA₄, LPA₅, LPA₆)with LPA mediates a range of downstream signaling cascades. The actualpathway and realized end point are dependent on a range of variablesthat include receptor usage, cell type, expression level of a receptoror signaling protein, and LPA concentration. Nearly all mammalian cells,tissues and organs co-express several LPA-receptor subtypes, whichindicates that LPA receptors signal in a cooperative manner LPA₁, LPA₂,and LPA₃ share high amino acid sequence similarity.

Illustrative Biological Activity

LPA regulates many important functions of fibroblasts in wound healing,including proliferation, migration, differentiation and contraction.Fibroblast proliferation is required in wound healing in order to fillan open wound. In contrast, fibrosis is characterized by intenseproliferation and accumulation of myofibroblasts that activelysynthesize ECM and proinflammatory cytokines. LPA can either increase orsuppress the proliferation of cell types important in wound healing.

Tissue injury initiates a complex series of host wound-healingresponses; if successful, these responses restore normal tissuestructure and function. If not, these responses can lead to tissuefibrosis and loss of function.

A number of muscular dystrophies are characterized by a progressiveweakness and wasting of musculature, and by extensive fibrosis. It hasbeen shown that LPA treatment of cultured myoblasts induced significantexpression of connective tissue growth factor (CTGF). CTGF subsequentlyinduces collagen, fibronectin and integrin expression and inducesdedifferentiation of these myoblasts. Treatment of a variety of celltypes with LPA induces reproducible and high level induction of CTGF.CTGF is a profibrotic cytokine, signaling down-stream and in parallelwith TGFβ.

LPA and LPA₁ play key pathogenic roles in pulmonary fibrosis. Fibroblastchemoattractant activity plays an important role in the lungs inpatients with pulmonary fibrosis. Profibrotic effects of LPA₁-receptorstimulation is explained by LPA₁-receptor-mediated vascular leakage andincreased fibroblast recruitment, both profibrotic events. The LPA-LPA₁pathway has a role in mediating fibroblast migration and vascularleakage in IPF. The end result is the aberrant healing process thatcharacterises this fibrotic condition.

The LPA-LPA2 pathway contributes to the activation of the TGF-β pathwayin pulmonary fibrosis. In some embodiments, compounds that inhibit LPA2show efficacy in the treatment of lung fibrosis. In some embodiments,compounds that inhibit both LPA1 and LPA2 show improved efficacy in thetreatment of lung fibrosis compared to compounds which inhibit only LPA1or LPA2.

LPA and LPA₁ are involved in the etiology of kidney fibrosis. In miceinvalidated for the LPA₁ receptor (LPA₁ (−/−), the development of renalfibrosis was significantly attenuated. Unilateral ureteral obstruction(UUO; animal model of renal fibrosis) mice treated with the LPA receptorantagonist Ki16425 closely resembled the LPA₁ (−/−) mice.

LPA is implicated in liver disease and fibrosis. Plasma LPA levels andserum autotoxin are elevated in hepatitis patients and animal models ofliver injury in correlation with increased fibrosis. LPA also regulatesliver cell function. LPA₁ and LPA₂ receptors are expressed by mousehepatic stellate cells and LPA stimulates migration of hepaticmyofibroblasts.

LPA is in involved in wound healing in the eye. LPA₁ and LPA₃ receptorsare detectable in the normal rabbit corneal epithelial cells,keratocytes and endothelial cells and LPA₁ and LPA₃ expression areincreased in corneal epithelial cells following injury.

LPA is present in the aqueous humor and the lacrimal gland fluid of therabbit eye and these levels are increased in a rabbit corneal injurymodel.

LPA induces actin stress fiber formation in rabbit corneal endothelialand epithelial cells and promotes contraction corneal fibroblasts. LPAalso stimulates proliferation of human retinal pigmented epithelialcells.

LPA is implicated in myocardial infarction and cardiac fibrosis. SerumLPA levels are increased in patients following mycocardial infarction(MI) and LPA stimulates proliferation and collagen production (fibrosis)by rat cardiac fibroblasts. Both LPA1 and LPA3 receptors are highlyexpressed in human heart tissue.

In one aspect, compounds of Formula (I) are used to treat or preventfibrosis in a mammal. In one aspect, compounds of Formula (I) are usedto treat or prevent fibrosis of an organ or tissue in a mammal.

The terms “fibrosis” or “fibrosing disorder,” as used herein, refers toconditions that are associated with the abnormal accumulation of cellsand/or fibronectin and/or collagen and/or increased fibroblastrecruitment and include but are not limited to fibrosis of individualorgans or tissues such as the heart, kidney, liver, joints, lung,pleural tissue, peritoneal tissue, skin, cornea, retina, musculoskeletaland digestive tract.

Exemplary diseases, disorders, or conditions that involve fibrosisinclude, but are not limited to: Lung diseases associated with fibrosis,e.g., idiopathic pulmonary fibrosis, pulmonary fibrosis secondary tosystemic inflammatory disease such as rheumatoid arthritis, scleroderma,lupus, cryptogenic fibrosing alveolitis, radiation induced fibrosis,chronic obstructive pulmonary disease (COPD), scleroderma, chronicasthma, silicosis, asbestos induced pulmonary or pleural fibrosis, acutelung injury and acute respiratory distress (including bacterialpneumonia induced, trauma induced, viral pneumonia induced, ventilatorinduced, non-pulmonary sepsis induced, and aspiration induced); Chronicnephropathies associated with injury/fibrosis (kidney fibrosis), e.g.,glomerulonephritis secondary to systemic inflammatory diseases such aslupus and scleroderma, diabetes, glomerular nephritis, focal segmentalglomerular sclerosis, IgA nephropathy, hypertension, allograft andAlport; Gut fibrosis, e.g., scleroderma, and radiation induced gutfibrosis; Liver fibrosis, e.g., cirrhosis, alcohol induced liverfibrosis, nonalcoholic steatohepatitis (NASH), biliary duct injury,primary biliary cirrhosis, infection or viral induced liver fibrosis(e.g., chronic HCV infection), and autoimmune hepatitis; Head and neckfibrosis, e.g., radiation induced; Corneal scarring, e.g., LASIK(laser-assisted in situ keratomileusis), corneal transplant, andtrabeculectomy; Hypertrophic scarring and keloids, e.g., burn induced orsurgical; and Other fibrotic diseases, e.g., sarcoidosis, scleroderma,spinal cord injury/fibrosis, myelofibrosis, vascular restenosis,atherosclerosis, arteriosclerosis, Wegener's granulomatosis, mixedconnective tissue disease, and Peyronie's disease.

In one aspect, a mammal suffering from one of the following non-limitingexemplary diseases, disorders, or conditions will benefit from therapywith a compound of Formula (I): atherosclerosis, thrombosis, heartdisease, vasculitis, formation of scar tissue, restenosis, phlobitis,COPD (chronic obstructive pulmonary disease), pulmonary hypertension,pulmonary fibrosis, pulmonary inflammation, bowel adhesions, bladderfibrosis and cystitis, fibrosis of the nasal passages, sinusitis,inflammation mediated by neutrophils, and fibrosis mediated byfibroblasts.

In one aspect, compounds of Formula (I) are used to treat adermatological disorders in a mammal Dermatological disorders include,but are not limited to, proliferative or inflammatory disorders of theskin such as, atopic dermatitis, bullous disorders, collagenoses,psoriasis, psoriatic lesions, dermatitis, contact dermatitis, eczema,urticaria, rosacea, wound healing, scarring, hypertrophic scarring,keloids, Kawasaki Disease, rosacea, Sjogren-Larsso Syndrome, urticaria.

In some embodiments, provided is a method of reducing lung injury,vascular leakage, inflammation and/or fibrosis in a mammal comprisingadministering to the mammal a selective LPA1 receptor antagonist. Insome embodiments, provided is a method of reducing lung injury, vascularleakage, inflammation and fibrosis in a mammal comprising administeringto the mammal a selective LPA1 receptor antagonist. In some embodiments,provided is a method of attenuating fibrosis in a mammal comprisingadministering a selective LPA1 receptor antagonist. In some embodiments,provided is a method of attenuating tissue remodeling and fibrosis in amammal comprising administering a selective LPA1 receptor antagonist.

In some embodiments, provided is a method of decreasing cytokineproduction in a mammal comprising administering a selective LPA1receptor antagonist. In some embodiments, the method of decreasingcytokine production in a mammal comprising administering a selectiveLPA1 receptor antagonist results in a reduction of tissue damage andfibrosis in a mammal.

In some embodiments, provided is a method of treating fibrosis is amammal comprising administering to the mammal a selective LPA1 receptorantagonist. In some embodiments, provided is a method of treatingfibrosis in a mammal while maintaining body weight in the mammalcomprising administering to the mammal a selective LPA1 receptorantagonist. In some embodiments, provided is a method of treatingrespiratory disease in a mammal comprising administering to the mammal aselective LPA1 receptor antagonist.

In some embodiments, provided is a method of treating fibrosis in amammal with a selective LPA1 receptor anatgonist, wherein the fibrosisin the mammal is not responsive to treatment with pirfenidone. In someembodiments, the LPA1 receptor antagonist is a compound of Formula (I).

As shown in the Examples, a selective LPA1 receptor antagonist reducedlung fibrosis, kidney fibrosis and liver fibrosis in various animalmodels of fibrosis.

LPA is released following tissue injury. LPA₁ plays a role in theinitiation of neuropathic pain. In one aspect, compounds of Formula (I)are used in the treatment of pain in a mammal. In one aspect, the painis acute pain or chronic pain. In another aspect, the pain isneuropathic pain. In another aspect, the pain is cancer pain. In oneaspect, compounds of Formula (I) are used in the treatment offibromyalgia.

Lysophospholipid receptor signaling plays a role in the etiology ofcancer. Lysophosphatidic acid (LPA) and its G protein-coupled receptors(GPCRs) LPA₁, LPA₂, and/or LPA₃ play a role in the development ofseveral types of cancers.

LPA contributes to tumorigenesis by increasing motility and invasivenessof cells. LPA has been implicated in the initiation or progression ofovarian cancer. LPA is present at significant concentrations (2-80 μM)in the ascitic fluid of ovarian cancer patients. LPA receptors (LPA2 andLPA3) are also overexpressed in ovarian cancer cells as compared tonormal ovarian surface epithelial cells. LPA has also been implicated inthe initiation or progression of prostate cancer, breast cancer,melanoma, head and neck cancer, bowel cancer (colorectal cancer),thyroid cancer, glioblastoma, and other cancers.

LPA receptors mediate both migration of and invasion by pancreaticcancer cell lines: Ki16425 and LPA₁-specific siRNA effectively blockedin vitro migration in response to LPA and peritoneal fluid (ascites)from pancreatic cancer patients; in addition, Ki16425 blocked theLPA-induced and ascites-induced invasion activity of a highly peritonealmetastatic pancreatic cancer cell line (Yamada et al, J. Biol. Chem.,279, 6595-6605, 2004).

Colorectal carcinoma cell lines show significant expression of LPA₁ mRNAand respond to LPA by cell migration and production of angiogenicfactors. Overexpression of LPA receptors has a role in the pathogenesisof thyroid cancer. LPA₃ was originally cloned from prostate cancercells, concordant with the ability of LPA to induce autocrineproliferation of prostate cancer cells.

LPA has stimulatory roles in cancer progression in many types of cancer.LPA is produced from and induces proliferation of prostate cancer celllines. LPA induces human colon carcinoma DLD1 cell proliferation,migration, adhesion, and secretion of angiogenic factors through LPA₁signalling. In other human colon carcinoma cells lines (HT29 and WiDR),LPA enhances cell proliferation and secretion of angiogenic factors. Inother colon cancer cell lines, LPA₂ and LPA₃ receptor activation resultsin proliferation of the cells. LPA₁ is implicated in bone metastasis andKi16425 has been shown to inhibit metastasis to bone in vivo (Boucharabaet al., Proc. Natl. Acad. Sci. USA, 103, 9643-9648, 2006).

In one aspect, a compound of Formula (I) is used in the treatment ofcancer. In one aspect, compounds of Formula (I) are used in thetreatment of malignant and benign proliferative disease. In one aspect,compounds of Formula (I) are used to prevent or reduce proliferation oftumor cells, invasion and metastasis of carcinomas, pleural mesotheliomaor peritoneal mesothelioma, cancer pain, bone metastases. In one aspectis a method of treating cancer in a mammal, the method comprisingadministering to the mammal a compound of Formula (I) and a secondtherapeutic agent, wherein the second therapeutic agent is ananti-cancer agent. In some embodiments, radiation therapy is also used.

The types of cancer include, but is not limited to, solid tumors (suchas those of the bladder, bowel, brain, breast, endometrium, heart,kidney, lung, lymphatic tissue (lymphoma), ovary, pancreas or otherendocrine organ (thyroid), prostate, skin (melanoma or basal cellcancer) or hematological tumors (such as the leukemias) at any stage ofthe disease with or without metastases.

In one aspect, LPA is a contributor to the pathogenesis of respiratorydiseases. Proinflammatory effects of LPA include degranulation of mastcells, contraction of smooth-muscle cells and release of cytokines fromdendritic cells. LPA induces the secretion of IL-8 from human bronchialepithelial cells. IL-8 is found in increased concentrations in BALfluids from patients with asthma, chronic obstructive lung disease,pulmonary sarcoidosis and acute respiratory distress syndrome and IL-8has been shown to exacerbate airway inflammation and airway remodelingof asthmatics. LPA1, LPA2 and LPA3 receptors have all been shown tocontribute to the LPA-induced IL-8 production.

Administration of LPA in vivo induces airway hyper-responsiveness,itch-scratch responses, infiltration and activation of eosinophils andneutrophils, vascular remodeling, and nociceptive flexor responses. LPAalso induces histamine release from mouse and rat mast cells. In oneaspect, the effects of LPA are mediated through LPA₁ and/or LPA₃. In oneaspect, compounds of Formula (I) are used in the treatment of variousallergic disorders in a mammal In one aspect, compounds of Formula (I)are used in the treatment of respiratory diseases, disorders orconditions in a mammal In one aspect, compounds of Formula (I) are usedin the treatment of asthma in a mammal In one aspect, compounds ofFormula (I) are used in the treatment of chronic asthma in a mammal.

The term “respiratory disease,” as used herein, refers to diseasesaffecting the organs that are involved in breathing, such as the nose,throat, larynx, eustachian tubes, trachea, bronchi, lungs, relatedmuscles (e.g., diaphragm and intercostals), and nerves. Respiratorydiseases include, but are not limited to, asthma, adult respiratorydistress syndrome and allergic (extrinsic) asthma, non-allergic(intrinsic) asthma, acute severe asthma, chronic asthma, clinicalasthma, nocturnal asthma, allergen-induced asthma, aspirin-sensitiveasthma, exercise-induced asthma, isocapnic hyperventilation, child-onsetasthma, adult-onset asthma, cough-variant asthma, occupational asthma,steroid-resistant asthma, seasonal asthma, seasonal allergic rhinitis,perennial allergic rhinitis, chronic obstructive pulmonary disease,including chronic bronchitis or emphysema, pulmonary hypertension,interstitial lung fibrosis and/or airway inflammation and cysticfibrosis, and hypoxia.

In one aspect, presented herein is the use of compounds of Formula (I)in the treatment or prevention of chronic obstructive pulmonary diseasein a mammal comprising administering to the mammal at least once aneffective amount of at least one compound of Formula (I). In addition,chronic obstructive pulmonary disease includes, but is not limited to,chronic bronchitis or emphysema, pulmonary hypertension, interstitiallung fibrosis and/or airway inflammation, and cystic fibrosis.

The nervous system is a major locus for LPA₁ expression. In one aspect,provided is a compound of Formula (I) for use in the treatment orprevention of a nervous system disorder in a mammal The term “nervoussystem disorder,” as used herein includes, but is not limited to,Alzheimer's Disease, cerebral edema, cerebral ischemia, stroke, multiplesclerosis, neuropathies, Parkinson's Disease, multiple sclerosis,retinal ischemia, post-surgical cognitive dysfunction, migraine,peripheral neuropathy/neuropathic pain, spinal cord injury, cerebraledema and head injury.

Angiogenesis, the formation of new capillary networks from pre-existingvasculature, is normally invoked in wound healing, tissue growth andmyocardial angiogenesis after ischemic injury. Peptide growth factorsand lysophospholipids control coordinated proliferation, migration,adhesion, differentiation and assembly of vascular endothelial cells(VECs) and surrounding vascular smooth-muscle cells (VSMCs). In oneaspect, dysregulation of the processes mediating angiogenesis leads toatherosclerosis, hypertension, tumor growth, rheumatoid arthritis anddiabetic retinopathy.

The specific effects of LPA are receptor-mediated.

In one aspect, compounds of Formula (I) are used to treat or preventcardiovascular disease in mammal, including but not limited to:arrhythmia (atrial or ventricular or both); atherosclerosis and itssequelae; angina; cardiac rhythm disturbances; myocardial ischemia;myocardial infarction; cardiac or vascular aneurysm; vasculitis, stroke;peripheral obstructive arteriopathy of a limb, an organ, or a tissue;reperfusion injury following ischemia of the brain, heart, kidney orother organ or tissue; endotoxic, surgical, or traumatic shock;hypertension, valvular heart disease, heart failure, abnormal bloodpressure; shock; vasoconstriction (including that associated withmigraines); vascular abnormality, inflammation, insufficiency limited toa single organ or tissue.

In one aspect, provided herein are methods for preventing or treatingvasoconstriction, atherosclerosis and its sequelae myocardial ischemia,myocardial infarction, aortic aneurysm, vasculitis and stroke comprisingadministering at least once to the mammal an effective amount of atleast one compound of Formula (I) or pharmaceutical composition ormedicament which includes a compound of Formula (I).

In one aspect, provided herein are methods for reducing cardiacreperfusion injury following myocardial ischemia and/or endotoxic shockcomprising administering at least once to the mammal an effective amountof at least one compound of Formula (I).

In one aspect, provided herein are methods for reducing the constrictionof blood vessels in a mammal comprising administering at least once tothe mammal an effective amount of at least one compound of Formula (I).

In one aspect, provided herein are methods for lowering or preventing anincrease in blood pressure of a mammal comprising administering at leastonce to the mammal an effective amount of at least one compound ofFormula (I).

LPA is associated with various inflammatory/immune diseases. In oneaspect, compounds of Formula (I) are used to treat or preventinflammation in a mammal. In one aspect, antagonists of LPA₁ and/or LPA₃find use in the treatment or prevention of inflammatory/immune disordersin a mammal.

Examples of inflammatory/immune disorders include psoriasis, rheumatoidarthritis, vasculitis, inflammatory bowel disease, dermatitis,osteoarthritis, asthma, inflammatory muscle disease, allergic rhinitis,vaginitis, interstitial cystitis, scleroderma, eczema, allogeneic orxenogeneic transplantation (organ, bone marrow, stem cells and othercells and tissues) graft rejection, graft-versus-host disease, lupuserythematosus, inflammatory disease, type I diabetes, pulmonaryfibrosis, dermatomyositis, Sjogren's syndrome, thyroiditis (e.g.,Hashimoto's and autoimmune thyroiditis), myasthenia gravis, autoimmunehemolytic anemia, multiple sclerosis, cystic fibrosis, chronic relapsinghepatitis, primary biliary cirrhosis, allergic conjunctivitis and atopicdermatitis.

Other Diseases, Disorders or Conditions

In accordance with one aspect, are methods for treating, preventing,reversing, halting or slowing the progression of LPA-dependent orLPA-mediated diseases or conditions once it becomes clinically evident,or treating the symptoms associated with or related to LPA-dependent orLPA-mediated diseases or conditions, by administering to the mammal acompound of Formula (I). In certain embodiments, the subject already hasa LPA-dependent or LPA-mediated disease or condition at the time ofadministration, or is at risk of developing a LPA-dependent orLPA-mediated disease or condition.

In certain aspects, are methods for preventing or treating eosinophiland/or basophil and/or dendritic cell and/or neutrophil and/or monocyteand/or T-cell recruitment comprising administering at least once to themammal an effective amount of at least one compound of Formula (I).

In certain aspects, are methods for the treatment of cystitis,including, e.g., interstitial cystitis, comprising administering atleast once to the mammal a therapeutically effective amount of at leastone compound of Formula (I).

In accordance with one aspect, methods described herein include thediagnosis or determination of whether or not a patient is suffering froma LPA-dependent or LPA-mediated disease or condition by administering tothe subject a therapeutically effective amount of a compound of Formula(I) and determining whether or not the patient responds to thetreatment.

In one aspect provided herein are compounds of Formula (I),pharmaceutically acceptable salts, pharmaceutically acceptable prodrugs,and pharmaceutically acceptable solvates thereof, which are antagonistsof at least one LPA receptor (e.g. LPA₁, LPA₂, LPA₃) and are used totreat patients suffering from one or more LPA-dependent or LPA-mediatedconditions or diseases, including, but not limited to, lung fibrosis,kidney fibrosis, liver fibrosis, scarring, asthma, rhinitis, chronicobstructive pulmonary disease, pulmonary hypertension, interstitial lungfibrosis, arthritis, allergy, psoriasis, inflammatory bowel disease,adult respiratory distress syndrome, myocardial infarction, aneurysm,stroke, cancer, pain, proliferative disorders and inflammatoryconditions. In some embodiments, LPA-dependent conditions or diseasesinclude those wherein an absolute or relative excess of LPA is presentand/or observed.

In any of the aforementioned aspects the LPA-dependent or LPA-mediateddiseases or conditions include, but are not limited to, organ fibrosis,asthma, allergic disorders, chronic obstructive pulmonary disease,pulmonary hypertension, lung or pleural fibrosis, peritoneal fibrosis,arthritis, allergy, cancer, cardiovascular disease, ult respiratorydistress syndrome, myocardial infarction, aneurysm, stroke, and cancer.

In one aspect, compounds of Formula (I) are used to improve the cornealsensitivity decrease caused by corneal operations such as laser-assistedin situ keratomileusis (LASIK) or cataract operation, cornealsensitivity decrease caused by corneal degeneration, and dry eye symptomcaused thereby.

In one aspect, presented herein is the use of compounds of Formula (I)in the treatment or prevention of ocular inflammation and allergicconjunctivitis, vernal keratoconjunctivitis, and papillaryconjunctivitis in a mammal comprising administering at least once to themammal an effective amount of at least one compound of Formula (I).

In one aspect, presented herein is the use of compounds of Formula (I)in the treatment or prevention of Sjogren disease or inflammatorydisease with dry eyes in a mammal comprising administering at least onceto the mammal an effective amount of at least one compound of Formula(I).

In one aspect, LPA and LPA receptors (e.g. LPA₁) are involved in thepathogenesis of osteoarthritis. In one aspect, presented herein is theuse of compounds of Formula (I) in the treatment or prevention ofosteoarthritis in a mammal comprising administering at least once to themammal an effective amount of at least one compound of Formula (I).

In one aspect, LPA receptors (e.g. LPA₁, LPA₃) contribute to thepathogenesis of rheumatoid arthritis. In one aspect, presented herein isthe use of compounds of Formula (I) in the treatment or prevention ofrheumatoid arthritis in a mammal comprising administering at least onceto the mammal an effective amount of at least one compound of Formula(I).

In one aspect, LPA receptors (e.g. LPA₁) contribute to adipogenesis. Inone aspect, presented herein is the use of compounds of Formula (I) inthe promotion of adipose tissue formation in a mammal comprisingadministering at least once to the mammal an effective amount of atleast one compound of Formula (I).

Compounds

In some embodiments, provided herein is a compound having the structureof Formula (I) or a pharmaceutically acceptable salt thereof:

wherein

-   -   R¹ is —CO₂H, —CO₂R^(D), —CN, tetrazolyl, —C(═O)NH₂, —C(═O)NHR¹⁰,        —C(═O)NHSO₂R¹⁰ or —C(═O)NHCH₂CH₂SO₃H; R^(D) is H or C₁-C₄alkyl;    -   L¹ is absent or C₁-C₆alkylene;    -   R³ is H, C₁-C₄alkyl, C₃-C₆cycloalkyl, or C₁-C₄-fluoroalkyl;    -   R⁷ is H or C₁-C₄alkyl;    -   R⁸ is H, C₁-C₄alkyl, or C₁-C₄-fluoroalkyl;    -   R¹⁰ is a C₁-C₆alkyl, C₁-C₆-fluoroalkyl, C₃-C₆cycloalkyl, or a        substituted or unsubstituted phenyl;    -   each of R^(A), R^(B), and R^(C) are independently selected from        H, F, Cl, Br, I, —CN, —OH, C₁-C₄alkyl, C₁-C₄-fluoroalkyl,        C₁-C₄-fluoroalkoxy, C₁-C₄alkoxy, and C₁-C₄heteroalkyl;    -   m is 0, 1, or 2; n is 0, 1, or 2; p is 0, 1, or 2.

For any and all of the embodiments, substituents are selected from amongfrom a subset of the listed alternatives. For example, in someembodiments, R¹ is —CO₂H, —CO₂R¹, —CN, tetrazolyl, or —C(═O)NHSO₂R¹⁰.For example, in some embodiments, R¹ is —CO₂H or —CO₂(R^(D)). In someembodiments, R^(D) is H, —CH₃, or —CH₂CH₃. In some embodiments, R¹ is—CO₂H. In some embodiments, R¹ is —C(═O)NHSO₂R¹⁰. In some embodiments,R¹ is a carboxylic acid bioisostere.

In some embodiments, R³ is C₁-C₄alkyl. In some embodiments, R³ is —CH₃.

In some embodiments, R⁷ is H.

In some embodiments, R⁸ is H, C₁-C₄alkyl, or C₁-C₄-fluoroalkyl. In someembodiments, R⁸ is H. In some embodiments, R⁸ is H or C₁-C₄alkyl. Insome embodiments, R⁸ is H. In some embodiments, R⁸ is H, —CH₃, or —CF₃.In some embodiments, R⁸ is —CH₃. In some embodiments, R⁸ is —CH₂CH₃.

In some embodiments, R¹⁰ is a C₁-C₆alkyl or a substituted orunsubstituted phenyl. In some embodiments, R¹⁰ is a C₁-C₆alkyl. In someembodiments, R¹⁰ is a substituted or unsubstituted phenyl.

In some embodiments, each R^(A) is independently selected from H, F, Cl,Br, I, —CH₃, —CF₃, —OH, —OCF₃, and —OCH₃. In some embodiments, eachR^(A) is independently selected from H, F, Cl, —CH₃, —CF₃, —OH, —OCF₃,and —OCH₃. In some embodiments, each R^(A) is independently selectedfrom H, F, Cl, —CH₃, —CF₃, and —OH. In some embodiments, each R^(A) isindependently selected from H, F, Cl, —CH₃, and —OH. In someembodiments, each R^(A) is H.

In some embodiments, each R^(B) is independently selected from H, F, Cl,Br, I, —CH₃, —CF₃, —OH, —OCF₃, and —OCH₃. In some embodiments, eachR^(B) is independently selected from H, F, Cl, —CH₃, —CF₃, and —OH. Insome embodiments, each R^(B) is independently selected from H, F, Cl,—CH₃, and —OH. In some embodiments, each R^(B) is H.

In some embodiments, each R^(C) is independently selected from H, F, Cl,Br, I, —CH₃, —CF₃, —OH, —OCF₃, and —OCH₃. In some embodiments, eachR^(C) is independently selected from H, F, Cl, —CH₃, —CF₃, —OH, —OCF₃,and —OCH₃. In some embodiments, each R^(C) is independently selectedfrom H, F, Cl, —CH₃, —CF₃, and —OH. In some embodiments, each R^(C) isindependently selected from H, F, Cl, and —OH. In some embodiments, eachR^(C) is independently selected from H, F, and Cl. In some embodiments,each R^(C) is H.

In some embodiments, m is 0 or 1. In some embodiments, m is 0. In someembodiments, m is 1. In some embodiments, p is 0 or 1. In someembodiments, p is 0. In some embodiments, p is 1. In some embodiments, nis 0, 1, or 2. In some embodiments, n is 0 or 1. In some embodiments, nis 0. In some embodiments, n is 1. In some embodiments, n is 2.

In some embodiments, R¹ is —CO₂H, —CO₂R^(D) or —C(═O)NHSO₂R¹⁰; R³ isC₁-C₄alkyl; R⁷ is H; R¹⁰ is a C₁-C₆alkyl or a substituted orunsubstituted phenyl; each R^(A) is independently selected from H, F,Cl, Br, I, —CH₃, —CF₃, —OH, —OCF₃, and —OCH₃; each R^(B) isindependently selected from H, F, Cl, Br, I, —CH₃, —CF₃, —OH, —OCF₃, and—OCH₃; each R^(C) is independently selected from H, F, Cl, Br, I, —CH₃,—CF₃, —OH, —OCF₃, and —OCH₃; m is 0 or 1; p is 0 or 1.

In some embodiments, R¹ is —CO₂H or —CO₂R¹; R³ is —CH₃ or —CH₂CH₃; R⁸ isH, —CH₃ or —CF₃; R^(D) is H, —CH₃, or —CH₂CH₃.

In some embodiments, R¹ is —CO₂H; R³ is —CH₃; R⁸ is —CH₃; L¹ is absent,—CH₂—, —CH₂CH₂—, —CH₂CH₂CH₂—, —CH(CH₃)—, —C(CH₃)₂—, —CH(CH₂CH₃)—,—C(CH₂CH₃)₂—, —CH₂CH(CH₃)—, or —CH₂C(CH₃)₂—. In some embodiments, R¹ is—CO₂H; R⁸ is —CH₃; L¹ is —CH₂—. In some embodiments, R¹ is —CO₂H; R⁸ is—CH₃; L¹ is —CH₂—; m is 0; n is 0 or 1; p is 0.

In some embodiments, the compound of Formula (I) has the followingstructure:

In some embodiments, L¹ is absent, —CH₂—, —CH(CH₃)—, —C(CH₃)₂—,—CH(CH₂CH₃)—, or —C(CH₂CH₃)₂—; each R^(C) is independently selected fromH, F, Cl, —CH₃, —CF₃, —OH, —OCF₃, and —OCH₃; m is 0; n is 0, 1, or 2; pis 0.

In some embodiments, L¹ is absent or —CH₂—; each R^(C) is independentlyselected from H, F, Cl, —CH₃, —CF₃, and —OH; n is 0 or 1.

In some embodiments, the compound of Formula (I) has the followingstructure:

In some embodiments, L¹ is absent, —CH₂—, —CH(CH₃)—, —C(CH₃)₂—,—CH(CH₂CH₃)—, or —C(CH₂CH₃)₂—; each R^(C) is independently selected fromH, F, Cl, —CH₃, —CF₃, —OH, —OCF₃, and —OCH₃; m is 0; n is 0, 1, or 2; pis 0.

In some embodiments, L¹ is absent or —CH₂—; each R^(C) is independentlyselected from H, F, Cl, —CH₃, —CF₃, and —OH; n is 0 or 1. In someembodiments, L¹ is —CH₂—.

In some embodiments, R¹ is —C(═O)NHSO₂R¹⁰; R³ is —CH₃ or —CH₂CH₃; R⁸ isH, —CH₃ or —CF₃; R¹⁰ is —CH₃, or —CH₂CH₃.

In some embodiments, L¹ is as described in Table 1 and/or Table 2.

In some embodiments,

as defined in Table 1.

In some embodiments,

as defined in Table 1.

In some embodiments,

In some embodiments,

In some embodiments, the compound of Formula (I) has a structureselected from:

Any combination of the groups described above for the various variablesis contemplated herein. Throughout the specification, groups andsubstituents thereof are chosen by one skilled in the field to providestable moieties and compounds.

In some embodiments, compounds of Formula (I) include, but are notlimited to, those described in Table 1, Table 2, Table 3 and FIGS. 1 to7.

TABLE 1

Cmpd # R^(A) B —L¹— R^(D) C R⁸ M + H* 1 H Phen-1,4- —CH₂— H2-Trifluoromethyl- CH₃ 525 ylene phenyl 2 H Phen-1,4- —CH₂— H3-Trifluoromethyl- CH₃ 525 ylene phenyl 3 H Phen-1,4- —CH₂— H2,4-Dichloro-phenyl CH₃ 525 ylene 4 H Phen-1,4- —CH₂— H 2-Fluoro-phenylCH₃ 475 ylene 5 H Phen-1,4- —CH₂— H 3-Bromo-phenyl CH₃ 535 ylene 6 HPhen-1,4- —CH₂— H 2-Methoxy-phenyl CH₃ 487 ylene 7 H 2-Methoxypen- —CH₂—H 2-Chloro-phenyl CH₃ 521 1,5-ylene 8 H Phen-1,4- — H 2-Chloro-phenylCH₃ 477 ylene 9 H Phen-1,2- — H 2-Chloro-phenyl CH₃ 477 ylene 10 HPhen-1,2- —CH₂— H 2-Chloro-phenyl CH₃ 491 ylene 11 H Phen-1,4- —CH₂— H2-Chloro-phenyl CH₃ 491 ylene 12 H Phen-1,3- —CH₂— H 2-Chloro-phenyl CH₃491 ylene 13 H Phen-1,4- —CH₂CH₃— H 2-Chloro-phenyl CH₃ 505 ylene 14 H2-Fluorophen- —CH₂— H 2-Chloro-phenyl CH₃ 509 1,5-ylene 15 H4-Fluorophen- —CH₂— H 2-Chloro-phenyl CH₃ 509 1,5-ylene 16 H Phen-1,4-—CH₂— —CH₃ 2-Chloro-phenyl CH₃ 505 ylene 17 H Phen-1,4- —CH(CH₃)——CH₂CH₃ 2-Chloro-phenyl CH₃ 533 ylene 18 H Phen-1,4- —CH(CH₃)— H2-Chloro-phenyl CH₃ 505 ylene 19 H Phen-1,4- —C(CH₃)₂— H 2-Chloro-phenylCH₃ 519 ylene 20 H Phen-1,4- —CH(CH₃)— H 2-Chloro-phenyl CH₃ 489 ylene21 H Phen-1,4- —CH₂CH₂CH₂— H 2-Chloro-phenyl CH₃ 519 ylene 22 HPhen-1,3- — H 2-Chloro-phenyl CH₃ 477 ylene 23 H Phen-1,4- —CH₂— H2-Fluoro-4- CH₃ 509 ylene chloro-phenyl 24 H Phen-1,4- —CH₂— H 2-Fluoro-(R)—CH₃ 475 ylene phenyl 25 —CH₃ Phen-1,4- —CH₂— H 2-Fluoro- (R)—CH₃ 489ylene phenyl 26 H Phen-1,4- —C(CH₃)₂— H 2-Fluoro- CH₃ 503 ylene phenyl27 H Phen-1,4- —CH₂— H 2-Chloro-phenyl (R)—CH₃ 491 ylene 28 H Phen-1,4-—C(CH₃)₂— H 2-Chloro-phenyl (R)—CH₃ 519 ylene 29 H Phen-1,4- —C(CH₃)₂— H2-Fluoro- (R)—CH₃ 503 ylene phenyl 30 H Phen-1,4- —CH(CH₃)— H 2-Fluoro-(R)—CH₃ 489 ylene phenyl 31 H Phen-1,4- —CH(CH₃)— H 2-Chloro-phenyl(R)—CH₃ 505 ylene 32 H Phen-1,4- —CH₂— H 2,6-Dichloro- CH₃ 525 ylenephenyl 33 —CH₃ Phen-1,4- —CH(CH₃)— H 2-Fluoro- (R)—CH₃ 503 ylene phenyl34 H Phen-1,4- —CH₂— H 2-Fluoro- (S)—CH₃ 475 ylene phenyl 35 H Phen-1,4-—CH₂— H 2-Chloro-phenyl (S)—CH₃ 491 ylene 36 H Phen-1,4- —CH₂— H2-Chloro-phenyl H 477 ylene 37 H Phen-1,4- —CH₂— H Phenyl (R)—CH₃ 457ylene 38 H Phen-1,4- —CH₂— H 2,3-Difluoro- CH₃ 493 ylene phenyl 39 HPhen-1,4- —CH₂— H 2,4-Difluoro- CH₃ 493 ylene phenyl 40 H Phen-1,4-—CH₂— H 2-Fluoro-4- CH₃ 505 ylene methoxy-phenyl 41 H Phen-1,4- —CH₂— H2,5-Difluoro- CH₃ 493 ylene phenyl 42 H Phen-1,4- —CH₂— H 2,6-Difluoro-CH₃ 493 ylene phenyl 43 H Phen-1,3- —CH₂— H Phenyl (R)—CH₃ 457 ylene 44H Phen-1,4- — H Phenyl (R)—CH₃ 443 ylene 45 H Phen-1,2- —CH₂— H Phenyl(R)—CH₃ 457 ylene 46 H Phen-1,4- —CH₂— H 2-Methyl-phenyl (R)—CH₃ 471ylene 47 H Phen-1,4- —CH(CH₃)— H 2-Chloro-phenyl (R)—CH₃ 505 ylene 48 HPhen-1,4- —CH(CH₃)— H 2-Chloro-phenyl (R)—CH₃ 505 ylene 49 H Phen-1,4-—CH₂— H 2-Chloro-phenyl (R)—CH₃ 525 ylene 50 H Phen-1,4- —CH(CH₂CH₃)— H2-Chloro-phenyl (R)—CH₃ 519 ylene 51 Cl Phen-1,4- —CH₂— H2-Chloro-phenyl (R)—CH₃ 526 ylene 52 F Phen-1,4- —CH₂— H 2-Chloro-phenyl(R)—CH₃ 509 ylene 53 H Phen-1,4- — H 2-Chloro-phenyl (R)—CH₃ 477 ylene56 H Phen-1,4- —CH₂— H 3,5-Dibromo-phenyl (R)—CH₃ 615 ylene 57 HPhen-1,4- —CH₂— H Phenyl (S)—CH₃ 457 ylene 58 H Phen-1,4- —CH₂— H3-Hydroxy-phenyl (R)—CH₃ 473 ylene 59 H Phen-1,4- —CH₂— H Phenyl —CH₃457 ylene 60 H Phen-1,4- —CH₂— H Phenyl-d5 —CD₃ 466 ylene *massspectrometric data

TABLE 2

Cmpd M + # L¹ R¹ R^(C) R⁸ H* 61 —CH₂— —CN Cl (R)—CH₃ 472 62 —CH₂— —CN F(R)—CH₃ 456 63 —CH₂— 2H-Tetrazol-5-yl F (R)—CH₃ 499 64 —CH₂—2H-Tetrazol-5-yl Cl (R)—CH₃ 515 65 —CH₂— —C(═NH)—NH₂ F (R)—CH₃ 473 66—CH₂— —C(═NH)—NH—Ac F (R)—CH₃ 515 67 —CH₂— —C(═O)—NHCH₂CH₂SO₃H H (R)—CH₃564 *mass spectrometric data

TABLE 3

Cmpd # R¹ R^(C) R⁸ 67 —C(═O)NHSO₂Me H (R)—CH₃ 68 —C(═O)NHSO₂Me H H 69—C(═O)NHSO₂Me 2-F (R)—CH₃ 70 —C(═O)NHSO₂Me 2-F H 71 —C(═O)NHSO₂Me 2-Cl(R)—CH₃ 72 —C(═O)NHSO₂Me 2-Cl H 73 —C(═O)NHSO₂Me 2-CH₃ (R)—CH₃ 74—C(═O)NHSO₂Me 2-CH₃ H 75 —C(═O)NHSO₂Me 2-CF₃ (R)—CH₃ 76 —C(═O)NHSO₂Me2-CF₃ H 77 —C(═O)NHSO₂Me 3-F (R)—CH₃ 78 —C(═O)NHSO₂Me 3-F H 79—C(═O)NHSO₂Me 3-Cl (R)—CH₃ 80 —C(═O)NHSO₂Me 3-Cl H 81 —C(═O)NHSO₂Me3-CH₃ (R)—CH₃ 82 —C(═O)NHSO₂Me 3-CH₃ H 83 —C(═O)NHSO₂Me 3-CF₃ (R)—CH₃ 84—C(═O)NHSO₂Me 3-CF₃ H 85 —C(═O)NHSO₂Ph H (R)—CH₃ 86 —C(═O)NHSO₂Ph H H 87—C(═O)NHSO₂Ph 2-F (R)—CH₃ 88 —C(═O)NHSO₂Ph 2-F H 89 —C(═O)NHSO₂Ph 2-Cl(R)—CH₃ 90 —C(═O)NHSO₂Ph 2-Cl H 91 —C(═O)NHSO₂Ph 2-CH₃ (R)—CH₃ 92—C(═O)NHSO₂Ph 2-CH₃ H 93 —C(═O)NHSO₂Ph 2-CF₃ (R)—CH₃ 94 —C(═O)NHSO₂Ph2-CF₃ H 95 —C(═O)NHSO₂Ph 3-F (R)—CH₃ 96 —C(═O)NHSO₂Ph 3-F H 97—C(═O)NHSO₂Ph 3-Cl (R)—CH₃ 98 —C(═O)NHSO₂Ph 3-Cl H 99 —C(═O)NHSO₂Ph3-CH₃ (R)—CH₃ 100 —C(═O)NHSO₂Ph 3-CH₃ H 101 —C(═O)NHSO₂Ph 3-CF₃ (R)—CH₃102 —C(═O)NHSO₂Ph 3-CF₃ HSynthesis of Compounds

Compounds of Formula (I) described herein are synthesized using standardsynthetic techniques or using methods known in the art in combinationwith methods described herein. In additions, solvents, temperatures andother reaction conditions presented herein may vary.

The starting material used for the synthesis of the compounds of Formula(I) are either synthesized or obtained from commercial sources, such as,but not limited to, Sigma-Aldrich, Fluka, Acros Organics, Alfa Aesar,and the like. General methods for the preparation of compounds can bemodified by the use of appropriate reagents and conditions for theintroduction of the various moieties found in the formulae as providedherein.

In some embodiments, the compounds of Formula (I) are prepared asoutlined herein.

Further Forms of Compounds

In one aspect, compounds of Formula (I) possess one or morestereocenters and each stereocenter exists independently in either the Ror S configuration. The compounds presented herein include alldiastereomeric, and enantiomeric forms. Stereoisomers are obtained, ifdesired, by methods such as, stereoselective synthesis and/or theseparation of stereoisomers by chiral chromatographic columns.

The methods and formulations described herein include the use ofN-oxides (if appropriate), crystalline forms (also known as polymorphs),amorphous phases, and/or pharmaceutically acceptable salts of compoundshaving the structure of Formula (I), as well as metabolites and activemetabolites of these compounds having the same type of activity. In somesituations, compounds may exist as tautomers. All tautomers are includedwithin the scope of the compounds presented herein. In specificembodiments, the compounds described herein exist in solvated forms withpharmaceutically acceptable solvents such as water, ethanol, and thelike. In other embodiments, the compounds described herein exist inunsolvated form.

In some embodiments, compounds described herein are prepared asprodrugs. A “prodrug” refers to an agent that is converted into theparent drug in vivo. In certain embodiments, upon in vivoadministration, a prodrug is chemically converted to the biologically,pharmaceutically or therapeutically active form of the compound. Incertain embodiments, a prodrug is enzymatically metabolized by one ormore steps or processes to the biologically, pharmaceutically ortherapeutically active form of the compound.

In some embodiments, sites on the aromatic ring portion of compounds ofFormula (I) are susceptible to various metabolic reactions.Incorporation of appropriate substituents on the aromatic ringstructures will reduce, minimize or eliminate this metabolic pathway. Inspecific embodiments, the appropriate substituent to decrease oreliminate the susceptibility of the aromatic ring to metabolic reactionsis, by way of example only, a deuterium, a halogen, or an alkyl group.

In another embodiment, the compounds described herein are labeledisotopically or by another other means, including, but not limited to,the use of chromophores or fluorescent moieties, bioluminescent labels,or chemiluminescent labels.

In one aspect, substitution with isotopes such as deuterium affordscertain therapeutic advantages resulting from greater metabolicstability, such as, for example, increased in vivo half-life or reduceddosage requirements.

“Pharmaceutically acceptable,” as used herein, refers a material, suchas a carrier or diluent, which does not abrogate the biological activityor properties of the compound, and is relatively nontoxic, i.e., thematerial may be administered to an individual without causingundesirable biological effects or interacting in a deleterious mannerwith any of the components of the composition in which it is contained.

In some embodiments, pharmaceutically acceptable salts are obtained byreacting a compound of Formula (I) with acids. Pharmaceuticallyacceptable salts are also obtained by reacting a compound of Formula (I)with a base to form a salt.

Compounds described herein may be formed as, and/or used as,pharmaceutically acceptable salts. The type of pharmaceutical acceptablesalts, include, but are not limited to: (1) acid addition salts, formedby reacting the free base form of the compound with a pharmaceuticallyacceptable inorganic acid (e.g. hydrochloric acid, hydrobromic acid,sulfuric acid, phosphoric acid, and the like); or with an organic acid(e.g. acetic acid, propionic acid, glycolic acid, pyruvic acid, lacticacid, malonic acid, succinic acid, malic acid, maleic acid, fumaricacid, trifluoroacetic acid, tartaric acid, citric acid, benzoic acid,cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid,1,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonicacid, toluenesulfonic acid, 2-naphthalenesulfonic acid, salicylic acid,stearic acid, muconic acid, butyric acid, phenylacetic acid,phenylbutyric acid, valproic acid, and the like); (2) salts formed whenan acidic proton present in the parent compound is replaced by a metalion, e.g., an alkali metal ion (e.g. lithium, sodium, potassium), analkaline earth ion (e.g. magnesium, or calcium), or an aluminum ion. Insome cases, compounds described herein may coordinate with an organicbase, such as, but not limited to, ethanolamine, diethanolamine,triethanolamine, tromethamine, N-methylglucamine, dicyclohexylamine,tris(hydroxymethyl)methylamine. In other cases, compounds describedherein may form salts with amino acids such as, but not limited to,arginine, lysine, and the like. Acceptable inorganic bases used to formsalts with compounds that include an acidic proton, include, but are notlimited to, aluminum hydroxide, calcium hydroxide, potassium hydroxide,sodium carbonate, sodium hydroxide, and the like. In some embodiments, asodium salt of the compound of Formula (I) is prepared.

It should be understood that a reference to a pharmaceuticallyacceptable salt includes the solvent addition forms or crystal formsthereof, particularly solvates or polymorphs. Solvates contain eitherstoichiometric or non-stoichiometric amounts of a solvent, and may beformed during the process of crystallization with pharmaceuticallyacceptable solvents such as water, ethanol, and the like. Hydrates areformed when the solvent is water, or alcoholates are formed when thesolvent is alcohol. In addition, the compounds provided herein can existin unsolvated as well as solvated forms. In general, the solvated formsare considered equivalent to the unsolvated forms for the purposes ofthe compounds and methods provided herein.

Compounds described herein, such as compounds of Formula (I), may be invarious forms, including but not limited to, amorphous forms, milledforms and nano-particulate forms. In addition, compounds describedherein include crystalline forms, also known as polymorphs. Polymorphsinclude the different crystal packing arrangements of the same elementalcomposition of a compound.

Certain Terminology

Unless otherwise stated, the following terms used in this application,including the specification and claims, have the definitions givenbelow. It must be noted that, as used in the specification and theappended claims, the singular forms “a,” “an” and “the” include pluralreferents unless the context clearly dictates otherwise. Unlessotherwise indicated, conventional methods of mass spectroscopy, NMR,HPLC, protein chemistry, biochemistry, recombinant DNA techniques andpharmacology are employed. In this application, the use of “or” or “and”means “and/or” unless stated otherwise. Furthermore, use of the term“including” as well as other forms, such as “include”, “includes,” and“included,” is not limiting. The section headings used herein are fororganizational purposes only and are not to be construed as limiting thesubject matter described.

An “alkyl” refers to an aliphatic hydrocarbon. The alkyl may besaturated or unsaturated. The alkyl, whether saturated or unsaturated,is a branched alkyl or straight chain alkyl. Typical alkyl groupsinclude, but are in no way limited to, methyl, ethyl, propyl, isopropyl,butyl, isobutyl, sec-butyl, tertiary butyl, pentyl, neopentyl, hexyl,allyl, but-2-enyl, but-3-enyl, and the like.

The term “alkylene” refers to a divalent alkyl radical. Typical alkylenegroups include, but are not limited to, —CH₂—, —CH(CH₃)—, —C(CH₃)₂—,—CH₂CH₂—, —CH₂CH(CH₃)—, —CH₂C(CH₃)₂—, —CH₂CH₂CH₂—, —CH₂CH₂CH₂CH₂—, andthe like.

An “alkoxy” group refers to a (alkyl)O— group, where alkyl is as definedherein.

“Cycloalkyl” refers to cyclopropyl, cyclobutyl, cyclopentyl,cyclopentenyl, cyclohexyl, and cyclohexenyl.

The term “halo” or, alternatively, “halogen” or “halide” means fluoro,chloro, bromo or iodo.

The term “fluoroalkyl” refers to an alkyl in which one or more hydrogenatoms are replaced by a fluorine atom.

The term “heteroalkyl” refers to an alkyl group in which one or moreskeletal atoms of the alkyl are selected from an atom other than carbon,e.g., oxygen, nitrogen (e.g. NH or Nalkyl), sulfur, or combinationsthereof. In some embodiments, one aspect, heteroalkyl refers to an alkylgroup in which one of the skeletal atoms of the alkyl is oxygen.

The term “optionally substituted” or “substituted” means that thereferenced group may be substituted with one or more additional group(s)individually and independently selected from halogen, —CN, —NH₂, —OH,—NH(CH₃), —N(CH₃)₂, —CO₂H, —CO₂alkyl, —C(═O)NH₂, —C(═O)NHalkyl,—C(═O)N(alkyl)₂, —S(═O)₂NH₂, —S(═O)₂NH(alkyl), —S(═O)₂N(alkyl)₂, alkyl,cycloalkyl, fluoroalkyl, heteroalkyl, alkoxy, fluoroalkoxy, —S-alkyl, or—S(═O)₂alkyl. In some embodiments, an optional substituent is selectedfrom halogen, —CN, —NH₂, —OH, —NH(CH₃), —N(CH₃)₂, —CH₃, —CH₂CH₃, —CF₃,—OCH₃, —OCH₂CH₃, and —OCF₃. In some embodiments, substituted groups aresubstituted with one or two of the preceding groups. In someembodiments, substituted groups are substituted with one of thepreceding groups.

The term “acceptable” with respect to a formulation, composition oringredient, as used herein, means having no persistent detrimentaleffect on the general health of the subject being treated.

The term “modulate,” as used herein, means to interact with a targeteither directly or indirectly so as to alter the activity of the target,including, by way of example only, to enhance the activity of thetarget, to inhibit the activity of the target, to limit the activity ofthe target, or to extend the activity of the target.

The term “modulator,” as used herein, refers to a molecule thatinteracts with a target either directly or indirectly. The interactionsinclude, but are not limited to, the interactions of an agonist, partialagonist, an inverse agonist and antagonist. In one embodiment, amodulator is an antagonist.

The term “agonist,” as used herein, refers to a molecule such as acompound, a drug, an enzyme activator or a hormone modulator that bindsto a specific receptor and triggers a response in the cell. An agonistmimics the action of an endogenous ligand (such as LPA, prostaglandin,hormone or neurotransmitter) that binds to the same receptor.

The term “antagonist,” as used herein, refers to a molecule such as acompound, which diminishes, inhibits, or prevents the action of anothermolecule or the activity of a receptor site. Antagonists include, butare not limited to, competitive antagonists, non-competitiveantagonists, uncompetitive antagonists, partial agonists and inverseagonists.

The term “LPA-dependent”, as used herein, refers to conditions ordisorders that would not occur, or would not occur to the same extent,in the absence of LPA.

The term “LPA-mediated”, as used herein, refers to refers to conditionsor disorders that might occur in the absence of LPA but can occur in thepresence of LPA.

“Selectivity” for one LPA receptor versus other LPA receptors means thatthe compound has an IC₅₀ (Ca Flux assay) for the indicated LPA receptorthat is at least 10-fold less than the IC₅₀ for other LPA receptors. Insome embodiments, selectivity for one LPA receptor versus other LPAreceptor means that the compound has an IC₅₀ for the indicated LPAreceptor that is at least 10-fold, at least 20-fold, at least 40-fold,at least 50-fold, at least 100-fold, at least 200-fold, at least500-fold, or at least 1000-fold, less than the IC₅₀ for other LPAreceptors. For example, a selective LPA₁ receptor antagonist has an IC₅₀that is at least 10-fold, at least 20-fold, at least 40-fold, at least50-fold, at least 100-fold, at least 200-fold, at least 500-fold, or atleast 1000-fold, less than the IC₅₀ for other LPA receptors (e.g. LPA₂,LPA₃).

The terms “co-administration” or the like, as used herein, are meant toencompass administration of the selected therapeutic agents to a singlepatient, and are intended to include treatment regimens in which theagents are administered by the same or different route of administrationor at the same or different time.

The terms “effective amount” or “therapeutically effective amount,” asused herein, refer to a sufficient amount of an agent or a compoundbeing administered which will relieve to some extent one or more of thesymptoms of the disease or condition being treated. The result can bereduction and/or alleviation of the signs, symptoms, or causes of adisease, or any other desired alteration of a biological system. Forexample, an “effective amount” for therapeutic uses is the amount of thecomposition comprising a compound as disclosed herein required toprovide a clinically significant decrease in disease symptoms. Anappropriate “effective” amount in any individual case may be determinedusing techniques, such as a dose escalation study.

The term “pharmaceutical combination” as used herein, means a productthat results from the mixing or combining of more than one activeingredient and includes both fixed and non-fixed combinations of theactive ingredients. The term “fixed combination” means that the activeingredients, e.g. a compound of Formula (I) and a co-agent, are bothadministered to a patient simultaneously in the form of a single entityor dosage. The term “non-fixed combination” means that the activeingredients, e.g. a compound of Formula (I) and a co-agent, areadministered to a patient as separate entities either simultaneously,concurrently or sequentially with no specific intervening time limits,wherein such administration provides effective levels of the twocompounds in the body of the patient. The latter also applies tococktail therapy, e.g. the administration of three or more activeingredients.

The term “subject” or “patient” encompasses mammals. Examples of mammalsinclude, but are not limited to, humans, chimpanzees, apes, monkey,cattle, horses, sheep, goats, swine, rabbits, dogs, cats, rodents, rats,mice guinea pigs, and the like. In one embodiment, the mammal is a human

The terms “treat,” “treating” or “treatment,” as used herein, includealleviating, abating or ameliorating at least one symptom of a diseaseor condition, preventing additional symptoms, inhibiting the disease orcondition, e.g., arresting the development of the disease or condition,relieving the disease or condition, causing regression of the disease orcondition, relieving a condition caused by the disease or condition, orstopping the symptoms of the disease or condition eitherprophylactically and/or therapeutically.

Pharmaceutical Compositions/Formulations and Routes of Administration

In some embodiments, the compounds described herein are formulated intopharmaceutical compositions. Pharmaceutical compositions are formulatedin a conventional manner using one or more pharmaceutically acceptableinactive ingredients that facilitate processing of the active compoundsinto preparations that can be used pharmaceutically. Proper formulationis dependent upon the route of administration chosen. A summary ofpharmaceutical compositions described herein can be found, for example,in Remington: The Science and Practice of Pharmacy, Nineteenth Ed(Easton, Pa.: Mack Publishing Company, 1995); Hoover, John E.,Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa.1975; Liberman, H. A. and Lachman, L, Eds., Pharmaceutical Dosage Forms,Marcel Decker, New York, N.Y., 1980; and Pharmaceutical Dosage Forms andDrug Delivery Systems, Seventh Ed. (Lippincott Williams & Wilkins 1999),herein incorporated by reference for such disclosure.

A pharmaceutical composition, as used herein, refers to a mixture of acompound of Formula (I) with other chemical components (i.e.pharmaceutically acceptable inactive ingredients), such as carriers,excipients, binders, filling agents, suspending agents, flavoringagents, sweetening agents, disintegrating agents, dispersing agents,surfactants, lubricants, colorants, diluents, solubilizers, moisteningagents, plasticizers, stabilizers, penetration enhancers, wettingagents, anti-foaming agents, antioxidants, preservatives, or one or morecombination thereof. The pharmaceutical composition facilitatesadministration of the compound to an organism.

Pharmaceutical formulations described herein are administrable to asubject in a variety of ways by multiple administration routes,including but not limited to, oral, parenteral (e.g., intravenous,subcutaneous, intramuscular, intramedullary injections, intrathecal,direct intraventricular, intraperitoneal, intralymphatic, intranasalinjections), intranasal, buccal, topical or transdermal administrationroutes. The pharmaceutical formulations described herein include, butare not limited to, aqueous liquid dispersions, self-emulsifyingdispersions, solid solutions, liposomal dispersions, aerosols, soliddosage forms, powders, immediate release formulations, controlledrelease formulations, fast melt formulations, tablets, capsules, pills,delayed release formulations, extended release formulations, pulsatilerelease formulations, multiparticulate formulations, and mixed immediateand controlled release formulations.

In some embodiments, the compounds of Formula (I) are administeredorally.

In some embodiments, the compounds of Formula (I) are administeredtopically. In such embodiments, the compound of Formula (I) isformulated into a variety of topically administrable compositions, suchas solutions, suspensions, lotions, gels, pastes, shampoos, scrubs,rubs, smears, medicated sticks, medicated bandages, balms, creams orointments. In one aspect, the compounds of Formula (I) are administeredtopically to the skin.

In another aspect, the compounds of Formula (I) are administered byinhalation.

In another aspect, the compounds of Formula (I) are formulated forintranasal administration. Such formulations include nasal sprays, nasalmists, and the like.

In another aspect, the compounds of Formula (I) are formulated as eyedrops.

In any of the aforementioned aspects are further embodiments in whichthe effective amount of the compound of Formula (I) is: (a) systemicallyadministered to the mammal; and/or (b) administered orally to themammal; and/or (c) intravenously administered to the mammal; and/or (d)administered by inhalation to the mammal; and/or (e) administered bynasal administration to the mammal; or and/or (f) administered byinjection to the mammal; and/or (g) administered topically to themammal; and/or (h) administered by ophthalmic administration; and/or (i)administered rectally to the mammal; and/or (j) adminsterednon-systemically or locally to the mammal

In any of the aforementioned aspects are further embodiments comprisingsingle administrations of the effective amount of the compound,including further embodiments in which (i) the compound is administeredonce; (ii) the compound is administered to the mammal multiple timesover the span of one day; (iii) continually; or (iv) continuously.

In any of the aforementioned aspects are further embodiments comprisingmultiple administrations of the effective amount of the compound,including further embodiments in which (i) the compound is administeredcontinuously or intermittently: as in a single dose; (ii) the timebetween multiple administrations is every 6 hours; (iii) the compound isadministered to the mammal every 8 hours; (iv) the compound isadministered to the mammal every 12 hours; (v) the compound isadministered to the mammal every 24 hours. In further or alternativeembodiments, the method comprises a drug holiday, wherein theadministration of the compound is temporarily suspended or the dose ofthe compound being administered is temporarily reduced; at the end ofthe drug holiday, dosing of the compound is resumed. In one embodiment,the length of the drug holiday varies from 2 days to 1 year.

In certain embodiments, a compound as described herein is administeredin a local rather than systemic manner

In some embodiments, the compound described herein is administeredtopically. In some embodiments, the compound described herein isadministered systemically.

In some embodiments, the pharmaceutical formulation is in the form of atablet. In other embodiments, pharmaceutical formulations of thecompounds of Formula (I) are in the form of a capsule.

In one aspect, liquid formulation dosage forms for oral administrationare in the form of aqueous suspensions or solutions selected from thegroup including, but not limited to, aqueous oral dispersions,emulsions, solutions, elixirs, gels, and syrups.

For administration by inhalation, a compound of Formula (I) isformulated for use as an aerosol, a mist or a powder.

For buccal or sublingual administration, the compositions may take theform of tablets, lozenges, or gels formulated in a conventional manner.

In some embodiments, compounds of Formula (I) are prepared astransdermal dosage forms.

In one aspect, a compound of Formula (I) is formulated into apharmaceutical composition suitable for intramuscular, subcutaneous, orintravenous injection.

In some embodiments, the compounds described herein may be administeredtopically and can be formulated into a variety of topicallyadministrable compositions, such as solutions, suspensions, lotions,gels, pastes, medicated sticks, balms, creams or ointments.

In some embodiments, the compounds of Formula (I) are formulated inrectal compositions such as enemas, rectal gels, rectal foams, rectalaerosols, suppositories, jelly suppositories, or retention enemas.

Methods of Dosing and Treatment Regimens

In one embodiment, the compounds of Formula (I) are used in thepreparation of medicaments for the treatment of LPA-dependent orLPA-mediated diseases or conditions. In addition, a method for treatingany of the diseases or conditions described herein in a subject in needof such treatment, involves administration of pharmaceuticalcompositions that include at least one compound of Formula (I) or apharmaceutically acceptable salt, active metabolite, prodrug, or solvatethereof, in therapeutically effective amounts to said subject.

In certain embodiments, the compositions containing the compound(s)described herein are administered for prophylactic and/or therapeutictreatments. In certain therapeutic applications, the compositions areadministered to a patient already suffering from a disease or condition,in an amount sufficient to cure or at least partially arrest at leastone of the symptoms of the disease or condition. Amounts effective forthis use depend on the severity and course of the disease or condition,previous therapy, the patient's health status, weight, and response tothe drugs, and the judgment of the treating physician. Therapeuticallyeffective amounts are optionally determined by methods including, butnot limited to, a dose escalation clinical trial.

In prophylactic applications, compositions containing the compoundsdescribed herein are administered to a patient susceptible to orotherwise at risk of a particular disease, disorder or condition.

In certain embodiments, the dose of drug being administered may betemporarily reduced or temporarily suspended for a certain length oftime (i.e., a “drug holiday”).

Doses employed for adult human treatment are typically in the range of0.01 mg-5000 mg per day or from about 1 mg to about 1000 mg per day. Inone embodiment, the desired dose is conveniently presented in a singledose or in divided doses.

Patient Selection

In any of the aforementioned aspects involving the prevention ortreatment of LPA-mediated diseases or conditions are further embodimentscomprising identifying patients by screening for LPA receptor gene SNPs.Patients can be further selected based on increased LPA receptorexpression in the tissue of interest. LPA receptor expression aredetermined by methods including, but not limited to, northern blotting,western blotting, quantitative PCR (qPCR), flow cytometry,autoradiography (using a small molecule radioligand or PET ligand). Insome embodiments, patients are selected based on the concentration ofserum or tissue LPA measured by mass spectrometry. In some embodiments,patients are selected based on a combination of the above markers(increased LPA concentrations and increased LPA receptor expression).

Combination Treatments

In certain instances, it is appropriate to administer at least onecompound of Formula (I) in combination with another therapeutic agent.

In one specific embodiment, a compound of Formula (I) is co-administeredwith a second therapeutic agent, wherein the compound of Formula (I) andthe second therapeutic agent modulate different aspects of the disease,disorder or condition being treated, thereby providing a greater overallbenefit than administration of either therapeutic agent alone.

For combination therapies described herein, dosages of theco-administered compounds vary depending on the type of co-drug(s)employed, on the specific drug(s) employed, on the disease or conditionbeing treated and so forth. In additional embodiments, whenco-administered with one or more other therapeutic agents, the compoundprovided herein is administered either simultaneously with the one ormore other therapeutic agents, or sequentially.

If administration is simultaneous, the multiple therapeutic agents are,by way of example only, provided in a single, unified form, or inmultiple forms.

In another embodiment described herein, methods for treatment ofproliferative disorders, including cancer, comprises administration to amammal a compound of Formula (I) in combination with one or moreanti-cancer agents and/or radiation therapy.

In one aspect, compounds of Formula (I) are to treat or reduce fibrosisin a mammal In one aspect, compounds of Formula (I) are administered incombination with one or more immunosuppresants. In some embodiments, acompound of Formula (I) is administered with corticosteroids.

In yet another embodiment described herein, methods for treatingLPA-dependent or LPA-mediated conditions or diseases, such as thetherapy of respiratory disorders (e.g., pulmonary fibrosis, asthma,COPD, rhinitis), comprises administration to a patient compounds,pharmaceutical compositions, or medicaments described herein incombination with at least one agent used in the treatment of respiratoryconditions.

In some embodiments, compounds of Formula (I) are administered to apatient in combination with anti-inflammatory agents.

In one embodiment, compounds of Formula (I) are administered to apatient in combination with inhaled corticosteroids.

EXAMPLES

These examples are provided for illustrative purposes only and not tolimit the scope of the claims provided herein.

Synthesis of Compounds

Example 1 Synthesis of (R)-2′-chloro-alpha-methylbenzyl alcohol

Using the procedure of Meier et al (Tetrahedron, 1996, 52, 589; Method3), 2′-chloroacetophenone (Aldrich) was reduced to give(R)-2′-chloro-alpha-methylbenzylalcohol in at least 80 e.e. % (HPLCanalysis of the acetate derivative (made by reacting the benzyl alcoholwith acetyl chloride and triethylamine in methylene chloride) usingChiralcel OD eluted with 99:1 Hexane:ethanol. R isomer retention time4.3 minutes).

Example 2 Synthesis of (S)-2′-chloro-alpha-methylbenzyl alcohol

Using the procedure of Meier et al (Tetrahedron, 1996, 52, 589; Method3), 2′-chloroacetophenone (Aldrich) was reduced to give(S)-2′-chloro-alpha-methylbenzylalcohol in at least 80 e.e. % (HPLCanalysis of the acetate derivative (made by reacting the benzyl alcoholwith acetyl chloride and triethylamine in methylene chloride) usingChiralcel OD eluted with 99:1 Hexane:ethanol. S isomer retention time5.3 minutes)

Example 3 Synthesis of (R)-2′-fluoro-alpha-methylbenzyl alcohol

Using the procedure of Meier et al (Tetrahedron, 1996, 52, 589; Method3), 2′-fluoroacetophenone (Aldrich) was reduced to give(R)-2′-fluoro-alpha-methylbenzylalcohol in at least 80 e.e. % (HPLCanalysis of the acetate derivative (made by reacting the benzyl alcoholwith acetyl chloride and triethylamine in methylene chloride) usingChiralcel OD eluted with 99.8:0.2 Hexane:ethanol. R isomer retentiontime 5.9 minutes).

Example 4 Synthesis of (S)-2′-fluoro-alpha-methylbenzyl alcohol

Using the procedure of Meier et al (Tetrahedron, 1996, 52, 589; Method3), 2′-fluoroacetophenone (Aldrich) was reduced to give(S)-2′-fluoro-alpha-methylbenzylalcohol in at least 80 e.e. % (HPLCanalysis of the acetate derivative (made by reacting the benzyl alcoholwith acetyl chloride and triethylamine in methylene chloride) usingChiralcel OD eluted with 99.8:0.2 Hexane:Ethanol. S isomer retentiontime 6.7 minutes).

Example 5 Step 1: Synthesis of (3-Bromo-4-methoxy-phenyl)-acetic acidethyl ester

To 3-bromo-4-methoxyphenylacetic acid (24 g, 97.9 mmol) in ethanol (240mL) was added thionyl chloride (7.8 mL, 107.7 mmol), and the reactionwas stirred at room temperature for 1 hour. Once no starting materialwas seen by analytical LCMS, the mixture was basified with saturatedaqueous NaHCO₃ and extracted with dichloromethane. The combined organiclayers were dried over MgSO₄, filtered, and concentrated to give thedesired product.

Step 2: Synthesis of[4-Methoxy-3-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-aceticacid ethyl ester

(3-Bromo-4-methoxy-phenyl)-acetic acid ethyl ester (27.4 g, 100.3 mmol),bis(pinacolato)diboron (25.47 g, 100.3 mmol), and potassium acetate(24.6 g, 250.8 mmol) were combined in 1,4-dioxane (250 mL) under N₂. Thesolution was purged with N₂, and then(1,1′-bis(diphenylphosphino)ferrocene)-dichloropalladium(II) (4.10 g,5.02 mmol) was added and the reaction was heated to 110° C. overnight.The mixture was filtered through Celite and partitioned between EtOAcand brine. The aqueous layer was separated and extracted twice withEtOAc, and the combined organic layers were dried and concentrated. Theresidue was purified by silica gel chromatography (20-60% EtOAc inhexanes) to give the title compound.

Example 6 Synthesis of5-(4-Bromo-phenyl)-3-methyl-isoxazole-4-carboxylic acid Step 1:3-Methylamino-but-2-enoic acid methyl ester

To a solution of methyl acetoacetate (29.4 g, 253 mmol) in methanol (30mL) was added methylamine (33 wt % in EtOH; 48 mL, 385 mmol) dropwise atroom temperature. The reaction was stirred for 1 hour, and thenconcentrated and dried to give the title compound as a white crystallinesolid.

Step 2: 2-(4-Bromo-benzoyl)-3-oxo-butyric acid methyl ester

To 3-methylamino-but-2-enoic acid methyl ester (5.0 g, 39.1 mmol) in THF(70 mL) was added pyridine (3.7 mL, 47 mmol) dropwise. The mixture wascooled to 0° C., and 4-bromobenzoyl chloride (8.55 g, 39.1 mmol) in THF(30 mL) was added dropwise. The reaction was stirred at room temperatureovernight, and then water was added. The mixture was extracted withEtOAc, and the combined organic layers were washed with water, dried,filtered, and concentrated to give the title compound.

Step 3: 5-(4-Bromo-phenyl)-3-methyl-isoxazole-4-carboxylic acid methylester

To a mixture of 2-(4-bromo-benzoyl)-3-oxo-butyric acid methyl ester (11g, 39 mmol) in acetic acid (50 mL) was added hydroxylamine hydrochloride(2.66 g, 39 mmol), and the reaction was stirred at 115° C. for 1 hours.After cooling, saturated aqueous NaHCO₃ was added to the mixture toadjust to pH 8. The solution was extracted with EtOAc, and the combinedorganic layers were washed with brine, dried, filtered, and concentratedto give the title compound.

Step 4: 5-(4-Bromo-phenyl)-3-methyl-isoxazole-4-carboxylic acid

Lithium hydroxide (2 g, 48 mmol) was added to a solution of5-(4-bromo-phenyl)-3-methyl-isoxazole-4-carboxylic acid methyl ester (39mmol) in methanol (50 mL) and water (10 mL), and the reaction wasstirred at 60° C. for 1 hour. Acidic work-up gave the title compound.

Example 7 Synthesis of[4-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic acidethyl ester and 4-((Ethoxycarbonyl)methyl)phenylboronic acid Step 1:[4-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic acidethyl ester

Ethyl 4-bromophenylacetate (12 g, 49 mmol), bis(pinacolato)diboron (12.4g, 59 mmol), 1,1′-bis(diphenylphosphino)ferrocene)-dichloropalladium(II)(2 g, 2.4 mmol), and potassium acetate (9.6 g, 98 mmol) were combined in1,4-dioxane (100 mL), and the reaction was heated to 80° C. for 6 hours.The mixture was worked-up to give the title compound.

Step 2: 4-((Ethoxycarbonyl)methyl)phenylboronic acid

[4-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic acidethyl (0.460 g, 1.58 mmol), sodium periodate (0.746 g, 3.49 mmol), andammonium acetate (0.309 g, 4.01 mmol) were combined in 1:1 acetone:waterand stirred overnight at room temperature. Aqueous work-up provided thetitle compound.

Example 8 Synthesis of[3-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic acidmethyl ester and 3-((Methoxycarbonyl)methyl)phenylboronic acid Step 1:[3-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic acidmethyl ester

Prepared according to the procedure described in Example 7, Step 1 usingmethyl 2-(3-bromophenyl)acetate and bis(pinacolato)diboron.

Step 2: 3-((Methoxycarbonyl)methyl)phenylboronic acid

Prepared according to the procedure described in Example 7, Step 2 using[3-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic acidmethyl ester.

Example 9 Synthesis of(4′-{3-Methyl-4-[1-(2-trifluoromethyl-phenyl)-ethoxycarbonylamino]-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid (Compound 1) Step 1:5-(4′-Ethoxycarbonylmethyl-biphenyl-4-yl)-3-methyl-isoxazole-4-carboxylicacid

5-(4-Bromo-phenyl)-3-methyl-isoxazole-4-carboxylic acid (2.0 g, 7.07mmol), [4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-aceticacid ethyl ester (2.46 g, 8.5 mmol),tetrakis(triphenylphosphine)palladium(0) (0.80 g, 0.70 mmol), and sodiumbicarbonate (4.3 g, 52 mmol) were combined in 1,4-dioxane (50 mL) andwater (10 mL), and the reaction was stirred overnight at 80° C. to givethe title compound.

Step 2:(4′-{3-Methyl-4-[1-(2-trifluoromethyl-phenyl)-ethoxycarbonylamino]-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid ethyl ester

5-(4′-Ethoxycarbonylmethyl-biphenyl-4-yl)-3-methyl-isoxazole-4-carboxylicacid (0.150 g, 0.41 mmol), alpha-methyl-2-trifluoromethylbenzyl alcohol(0.094 g, 0.49 mmol), diphenylphosphoryl azide (0.11 mL, 0.49 mmol), andtriethylamine (0.11 mL, 0.49 mmol) were combined in toluene (3 mL) andstirred at 80° C. for 2 hours. After cooling, the mixture was worked-upto give the title compound.

Step 3:(4′-{3-Methyl-4-[1-(2-trifluoromethyl-phenyl)-ethoxycarbonylamino]-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid

(4′-{3-Methyl-4-[1-(2-trifluoromethyl-phenyl)-ethoxycarbonylamino]-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid ethyl ester (0.41 mmol) was treated with aqueous lithium hydroxidein methanol at 60° C. for 30 minutes. The mixture was purified bypreparative HPLC to give the title compound.

Example 10 Synthesis of(4′-{3-Methyl-4-[1-(3-trifluoromethyl-phenyl)-ethoxycarbonylamino]-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid (Compound 2)

Following the procedure described in Example 9, Step 2,5-(4′-ethoxycarbonylmethyl-biphenyl-4-yl)-3-methyl-isoxazole-4-carboxylicacid and alpha-methyl-3-trifluoromethylbenzyl alcohol were reacted toprovide(4′-{3-methyl-4-[1-(3-trifluoromethyl-phenyl)-ethoxycarbonylamino]-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid ethyl ester, which was hydrolyzed to the acid as described inExample 9, Step 3.

Example 11 Synthesis of(4′-{4-[1-(2,4-Dichloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid (Compound 3)

Following the procedure described in Example 9, Step 2,5-(4′-ethoxycarbonylmethyl-biphenyl-4-yl)-3-methyl-isoxazole-4-carboxylicacid and 2,4-dichloro-alpha-methylbenzyl alcohol were reacted to provide(4′-{4-[1-(2,4-Dichloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid ethyl ester, which was hydrolyzed to the acid as described inExample 9, Step 3.

Example 12 Synthesis of(4′-{4-[1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid (Compound 4)

Following the procedure described in Example 9, Step 2,5-(4′-ethoxycarbonylmethyl-biphenyl-4-yl)-3-methyl-isoxazole-4-carboxylicacid and 2-fluoro-alpha-methylbenzyl alcohol were reacted to provide(4′-{4-[1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid ethyl ester, which was hydrolyzed to the acid as described inExample 9, Step 3.

Example 13 Synthesis of(4′-{4-[1-(3-Bromo-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid (Compound 5)

Following the procedure described in Example 9, Step 2,5-(4′-ethoxycarbonylmethyl-biphenyl-4-yl)-3-methyl-isoxazole-4-carboxylicacid and 3-bromo-alpha-methylbenzyl alcohol were reacted to provide(4′-{4-[1-(3-bromo-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid ethyl ester, which was hydrolyzed to the acid as described inExample 9, Step 3.

Example 14 Synthesis of(4′-{4-[1-(2-Methoxy-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid (Compound 6)

Following the procedure described in Example 9, Step 2,5-(4′-ethoxycarbonylmethyl-biphenyl-4-yl)-3-methyl-isoxazole-4-carboxylicacid and 1-(2-methoxyphenyl)ethanol were reacted to provide(4′-{4-[1-(2-Methoxy-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid ethyl ester, which was hydrolyzed to the acid as described inExample 9, Step 3.

Example 15 Synthesis of (3-Bromo-4-fluoro-phenyl)-acetic acid ethylester

Step 1: 3-Bromo-4-fluorophenylacetic acid (2.35 g, 9.0 mmol) was treatedwith concentrated sulfuric acid (2 mL) in ethanol (100 mL) at reflux for2 hours. Purification by silica gel chromatography provided the titlecompound.

Example 16 Synthesis of (5-Bromo-2-fluoro-phenyl)-acetic acid ethylester

Step 1: Prepared according to the procedure described in Example 15,Step 1 using the following starting material:5-bromo-2-fluorophenylacetic acid.

Example 17 Synthesis of(4′-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-6-methoxy-biphenyl-3-yl)-aceticacid (Compound 7)

Step 1: To 5-(4-bromo-phenyl)-3-methyl-isoxazole-4-carboxylic acid(5.764 g, 20.4 mmol) in toluene (200 mL) was added triethylamine (3.13mL, 22.4 mmol) and diphenylphosphoryl azide (4.42 mL, 20.4 mmol), andthe mixture was stirred for 5 minutes at room temperature.2-Chloro-alpha-methylbenzyl alcohol (3.2 g, 22.4 mol) was added, and thereaction was stirred at 80° C. for 2 hours. The mixture was diluted withEtOAc (200 mL) and extracted with EtOAc. The combined organic layerswere washed twice with water and once with brine, and the combinedaqueous layers were back-extracted with EtOAc. The combined organiclayers were concentrated, and the residue was purified by silica gelchromatography to give[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid1-(2-chloro-phenyl)-ethyl ester.

Step 2: To [5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid1-(2-chloro-phenyl)-ethyl ester (0.100 g, 0.23 mmol) in DME (5 mL) wasadded[4-methoxy-3-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-aceticacid ethyl ester (0.074 g, 0.23 mmol), followed by potassium carbonate(0.079 g, 0.58 mmol) and water (3 mL). The solution was purged with N₂for 10 minutes, and then tetrakis(triphenylphosphine)palladium(0) (0.027g, 0.02 mmol) was added. The reaction was stirred at 100° C. until nostarting material was seen by analytical LCMS and thin layerchromatography (tlc), and then cooled to room temperature and dilutedwith EtOAc. The mixture was extracted with EtOAc, and the combinedorganic layers were washed with water and brine. The combined aqueouslayers were back-extracted with EtOAc, and the combined organic layerswere dried and concentrated. The residue was purified by silica gelchromatography to give(4′-{4-[1-(2-chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-6-methoxy-biphenyl-3-yl)-aceticacid ethyl ester.

Step 3: To(4′-{4-[1-(2-chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-6-methoxy-biphenyl-3-yl)-aceticacid ethyl ester (0.105 g, 0.20 mmol) in methanol (20 mL) was added 1Naqueous LiOH (5 mL, 5 mmol), and the reaction was stirred for 2 hours.The mixture was diluted with water and extracted with EtOAc. Thecombined organic layers were dried and concentrated to give the titlecompound.

Example 18 Synthesis of{3-Methyl-5-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-isoxazol-4-yl}-carbamicacid 1-(2-chloro-phenyl)-ethyl ester

[5-(4-Bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid1-(2-chloro-phenyl)-ethyl ester (3.48 g, 8.0 mmol),bis(pinacolato)diboron (2.24 g, 8.8 mmol), and potassium acetate (2.88g, 32.0 mmol) were combined in 1,4-dioxane (200 mL) under N₂. Thesolution was purged with N₂ for 10 minutes, and then(1,1′-bis(diphenylphosphino)ferrocene)-dichloropalladium(II) (0.652 g,0.8 mmol) was added and the reaction was heated to 80° C. for 6 hours,and then cooled to room temperature and stirred overnight. The mixturewas diluted with water and extracted with EtOAc. The combined organiclayers were washed twice with water and once with brine, and thecombined aqueous layers were back-extracted with EtOAc. The combinedorganic layers were concentrated and the residue was purified by silicagel chromatography (0-100% EtOAc in hexanes) to give the title compound.

Example 19 Synthesis of4′-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-carboxylicacid (Compound 8)

Step 1:{3-Methyl-5-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-isoxazol-4-yl}-carbamicacid 1-(2-chloro-phenyl)-ethyl ester (0.100 g, 0.21 mmol),4-bromobenzoic acid (0.042 g, 0.21 mmol), and potassium carbonate (0.073g, 0.53 mmol) were combined in DME (10 mL) and water (5 mL). Thesolution was purged with N₂ for 10 minutes, and then(1,1′-bis(diphenylphosphino)ferrocene)-dichloropalladium(II) (0.014 g,0.02 mmol) was added. The reaction was stirred at 70° C. overnight,until no starting material was seen by analytical LCMS. The mixture wascooled to room temperature and quenched with water. The mixture wasextracted with EtOAc, and the combined organic layers were dried andconcentrated. The residue was purified by preparative HPLC to give thetitle compound.

Example 20 Synthesis of4′-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-2-carboxylicacid (Compound 9)

Step 1:{3-Methyl-5-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-isoxazol-4-yl}-carbamicacid 1-(2-chloro-phenyl)-ethyl ester, methyl 2-bromobenzoate, andbis(triphenylphosphine)palladium(II) dichloride were reacted asdescribed in Example 17, Step 2 to provide4′-{4-[1-(2-chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-2-carboxylicacid methyl ester.

Step 2: To4′-{4-[1-(2-chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-2-carboxylicacid methyl ester (0.2 μmol) in methanol was added 1N aqueous LiOH (2mL) and stirred overnight at room temperature. The solution was stirredat 40° C. for 3 days. The mixture was purified by preparative HPLC togive the title compound.

Example 21 Synthesis of(4′-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-2-yl)-aceticacid (Compound 10)

Step 1: Prepared according to the procedure described in Example 19,Step 1 using{3-methyl-5-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-isoxazol-4-yl}-carbamicacid 1-(2-chloro-phenyl)-ethyl ester and 2-bromophenylacetic acid.

Example 22 Synthesis of(4′-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid (Compound 11)

Following the procedure described in Example 17, Step 2,{3-methyl-5-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-isoxazol-4-yl}-carbamicacid 1-(2-chloro-phenyl)-ethyl ester, ethyl 4-bromophenylacetate, and(1,1′-bis(diphenylphosphino)ferrocene)-dichloropalladium(II) werereacted to provide(4′-{4-[1-(2-chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid ethyl ester, which was hydrolyzed to the acid as described inExample 17, Step 3.

Example 23 Synthesis of(4′-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-3-yl)-aceticacid (Compound 12)

3-Bromophenylacetic acid (0.31 mmol) was treated with concentratedsulfuric acid in ethanol at reflux for 2 hours to give(3-bromo-phenyl)-acetic acid ethyl ester.

Following the procedure described in Example 17, Step 2,{3-methyl-5-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-isoxazol-4-yl}-carbamicacid 1-(2-chloro-phenyl)-ethyl ester, (3-bromo-phenyl)-acetic acid ethylester, and (1,1′-bis(diphenylphosphino)ferrocene)-dichloropalladium(II)were reacted to provide(4′-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-3-yl)-aceticacid ethyl ester, which was hydrolyzed to the acid as described inExample 17, Step 3.

Example 24 Synthesis of3-(4′-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-propionicacid (Compound 13)

3-(4-Bromo-phenyl)-propionic acid ethyl ester was prepared according tothe procedure described in Example 15, Step 1 using3-(4-bromophenyl)propionic acid.

Following the procedure described in Example 17, Step 2,{3-methyl-5-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-isoxazol-4-yl}-carbamicacid 1-(2-chloro-phenyl)-ethyl ester, 3-(4-bromo-phenyl)-propionic acidethyl ester, and bis(triphenylphosphine)palladium(II) dichloride werereacted to provide3-(4′-{4-[1-(2-chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-propionicacid ethyl ester, which was hydrolyzed to the acid as described inExample 17, Step 3.

Example 25 Synthesis of(4′-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-6-fluoro-biphenyl-3-yl)-aceticacid (Compound 14)

Following the procedure described in Example 17, Step 2,{3-methyl-5-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-isoxazol-4-yl}-carbamicacid 1-(2-chloro-phenyl)-ethyl ester, (3-bromo-4-fluoro-phenyl)-aceticacid ethyl ester, and(1,1′-bis(diphenylphosphino)ferrocene)-dichloropalladium(II) werereacted to provide(4′-{4-[1-(2-chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-6-fluoro-biphenyl-3-yl)-aceticacid ethyl ester, which was hydrolyzed to the acid as described inExample 17, Step 3.

Example 26 Synthesis of(4′-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-4-fluoro-biphenyl-3-yl)-aceticacid (Compound 15)

Following the procedure described in Example 17, Step 2,{3-methyl-5-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-isoxazol-4-yl}-carbamicacid 1-(2-chloro-phenyl)-ethyl ester, (5-bromo-2-fluoro-phenyl)-aceticacid ethyl ester, and(1,1′-bis(diphenylphosphino)ferrocene)-dichloropalladium(II) werereacted to provide(4′-{4-[1-(2-chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-4-fluoro-biphenyl-3-yl)-aceticacid ethyl ester, which was hydrolyzed to the acid as described inExample 17, Step 3.

Example 27 Synthesis of(4′-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid methyl ester (Compound 16)

To(4′-{4-[1-(2-chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid (0.050 g, 0.1 mmol) in benzene (17 mL) and methanol (2 mL) wasadded (trimethylsilyl)diazomethane (2M in hexanes; 0.1 mL, 0.2 mmol),and the reaction was stirred at room temperature for 5 minutes.Additional (trimethylsilyl)diazomethane (2M in hexanes; 0.25 mL, 0.5mmol) was added, and the reaction was stirred until no starting materialwas seen by analytical LCMS. The mixture was quenched with water andextracted with EtOAc. The combined organic layers were washed twice withwater and once with brine, and then dried and concentrated. The residuewas purified by silica gel chromatography (0-80% EtOAc in hexanes) togive the title compound.

Example 28 Synthesis of2-(4′-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-propionicacid ethyl ester (Compound 17)

Step 1: To ethyl 4-bromophenylacetate (2.137 g, 8.8 mmol) in THF (20 mL)at −78° C. was added lithium bis(trimethylsilyl)amide (1M in hexane;11.4 mL, 11.4 mmol), and the solution was stirred for 1 hour at −78° C.Iodomethane (0.71 mL, 11.4 mmol) in THF (5 mL) was added, and thereaction was warmed to room temperature. The mixture was quenched withwater and concentrated. The residue was dissolved in EtOAc and washedthree times with water and once with brine. The organic layer was driedand concentrated, and the crude material was purified by silica gelchromatography (0-60% EtOAc in hexanes) to give2-(4-bromo-phenyl)-propionic acid ethyl ester.

Step 2: Following the procedure described in Example 17, Step 2 thetitle compound was prepared using{3-methyl-5-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-isoxazol-4-yl}-carbamicacid 1-(2-chloro-phenyl)-ethyl ester, 2-(4-bromo-phenyl)-propionic acidethyl ester, and bis(triphenylphosphine)palladium(II) dichloride;further purification by preparative HPLC was required.

Example 29 Synthesis of2-(4′-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-propionicacid (Compound 18)

Prepared according to the procedure described in Example 17, Step 3using2-(4′-{4-[1-(2-chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-propionicacid ethyl ester.

Example 30 Synthesis of2-(4′-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-2-methyl-propionicacid (Compound 19)

To sodium hydride (0.859 g, 35.8 mmol) and iodomethane (2.23 mL, 35.8mmol) in dimethylformamide (80 mL) at 0° C. was added ethyl4-bromophenylacetate (2.17 g, 9.0 mmol) dropwise, and the reaction waswarmed to room temperature and stirred for 4 hours. The mixture wasquenched with water and 1N aqueous HCl (20 mL), and then extracted with1:1 EtOAc:hexanes. The combined organic layers were washed five timeswith water and once with brine, and the combined aqueous layers wereback-extracted with EtOAc. The combined organic layers were dried andconcentrated, and the residue was purified by silica gel chromatography(0-20% EtOAc in hexanes). Further purification by preparative HPLCprovided 2-(4-bromo-phenyl)-2-methyl-propionic acid ethyl ester.

Following the procedure described in Example 17, Step 2,{3-methyl-5-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-isoxazol-4-yl}-carbamicacid 1-(2-chloro-phenyl)-ethyl ester,2-(4-bromo-phenyl)-2-methyl-propionic acid ethyl ester, andbis(triphenylphosphine)palladium(II) dichloride were reacted to provide2-(4′-{4-[1-(2-chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-2-methyl-propionicacid ethyl ester; further purification by preparative HPLC was required.The ester was hydrolyzed to the acid as described in Example 17, Step 3;purification by preparative HPLC was utilized.

Example 31 Synthesis of2-(4′-{4-[1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-propionicacid (Compound 20)

Step 1: 5-(4-Bromo-phenyl)-3-methyl-isoxazole-4-carboxylic acid (4 g,14.2 mmol) was dissolved in toluene (150 mL). Triethylamine (2.187 mL,15.6 mmol) was added, followed by diphenylphosphoryl azide (3.372 mL,15.6 mmol), and the mixture was stirred for 10 minutes.2-Fluoro-alpha-methylbenzyl alcohol (2 g, 15.6 mmol) was added, and thereaction was stirred at 80° C. overnight. The mixture was cooled to roomtemperature and concentrated, and the residue was partitioned betweenwith EtOAc and water. The organic layer was washed 4 times with waterand once with brine, and then dried, filtered, and concentrated, and theresidue was purified by silica gel chromatography (dry load; 0-100%EtOAc in hexanes) to give[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid1-(2-fluoro-phenyl)-ethyl ester.

Step 2:{3-Methyl-5-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-isoxazol-4-yl}-carbamicacid 1-(2-fluoro-phenyl)-ethyl ester

Prepared according to the procedure described in Example 18 using[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid1-(2-fluoro-phenyl)-ethyl ester and bis(pinacolato)diboron.

Step 3:2-(4′-{4-[1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-propionicacid ethyl ester

Prepared according to the procedure described in Example 17, Step 2using{3-methyl-5-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-isoxazol-4-yl}-carbamicacid 1-(2-fluoro-phenyl)-ethyl ester, 2-(4-bromo-phenyl)-propionic acidethyl ester, and bis(triphenylphosphine)palladium(II) dichloride;purification by preparative HPLC, followed by silica gel chromatography,was utilized.

Step 4:2-(4′-{4-[1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-propionicacid

Prepared as described in Example 17, Step 3 using2-(4′-{4-[1-(2-fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-propionicacid ethyl ester.

Example 32 Synthesis of4-(4′-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-butyricacid (Compound 21)

4-(4-Bromo-phenyl)-butyric acid ethyl ester was prepared according tothe procedure described in Example 23 using 4-(4-bromophenyl)butanoicacid.

Following the procedure described in Example 17, Step 2,{3-methyl-5-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-isoxazol-4-yl}-carbamicacid 1-(2-chloro-phenyl)-ethyl ester, 4-(4-bromo-phenyl)-butyric acidethyl ester, and bis(triphenylphosphine)palladium(II) dichloride werereacted to provide4-(4′-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-butyricacid ethyl ester, which was hydrolyzed to the acid as described inExample 17, Step 3.

Example 33 Synthesis of4′-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-3-carboxylicacid (Compound 22)

Step 1: [5-(4-Bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid1-(2-chloro-phenyl)-ethyl ester (0.200 g, 0.46 mmol),3-carboxyphenylboronic acid (0.092 g, 0.55 mmol), and potassiumcarbonate (0.190 g, 1.37 mmol) were combined in 2:1 DME:water (5 mL),and the solution was purged with N₂ for 10 minutes.Tetrakis(triphenylphosphine)palladium(0) (0.053 g, 0.05 mmol) was added,and the reaction was purged with N₂ for an additional 10 minutes, andthen sealed and stirred at 80° C. overnight. The mixture was partitionedbetween EtOAc and water, and the aqueous layer was extracted with EtOAc.The combined organic layers were dried over MgSO₄, filtered, andconcentrated, and the residue was purified by preparative HPLC to givethe title compound.

Example 34 Synthesis of(4′-{4-[1-(4-Chloro-2-fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid (Compound 23)

Step 1:5-(4′-Ethoxycarbonylmethyl-biphenyl-4-yl)-3-methyl-isoxazole-4-carboxylicacid and 1-(4-chloro-2-fluorophenyl)ethanol were reacted as described inExample 31, Step 1 to provide(4′-{4-[1-(4-Chloro-2-fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid ethyl ester.

Step 2: Excess lithium hydroxide was added to a solution of(4′-{4-[1-(4-chloro-2-fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid ethyl ester (0.100 g, 0.19 mmol) in methanol and water, and thereaction was stirred at room temperature overnight. The mixture wasacidified with 1N aqueous HCl and extracted with EtOAc, and the combinedorganic layers were dried, filtered, and concentrated. The residue waspurified by preparative HPLC to give the title compound.

Example 35 Synthesis of(4′-{4-[(R)-1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid (Compound 24)

Step 1: 2′-Fluoroacetophenone (25 g, 180 mmol) and(S)-(−)-2-methyl-CBS-oxazaborolidine (6.02 g, 22 mmol) were combined inTHF (200 mL). Borane methyl sulfide complex (2M in THF; 59.4 mL, 119mmol) was added over 20 minutes, and the reaction was stirred at roomtemperature for 30 minutes. Methanol was added, and the mixture wasworked-up with dichloromethane and water. The organic layer wasconcentrated to give (R)-1-(2-fluoro-phenyl)-ethanol in 90.6% e.e.

Step 2:5-(4′-Ethoxycarbonylmethyl-biphenyl-4-yl)-3-methyl-isoxazole-4-carboxylicacid and (R)-1-(2-fluoro-phenyl)-ethanol were reacted as described inExample 31, Step 1 to provide(4′-{4-[(R)-1-(2-fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid ethyl ester.

Step 3: The title compound was prepared as described in Example 34, Step2 using(4′-{4-[(R)-1-(2-fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid ethyl ester.

Example 36 Synthesis of(4′-{4-[(R)-1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-2′-methyl-biphenyl-4-yl)-aceticacid (Compound 25) Step 1: 4-Bromo-3-methyl-benzoyl chloride

To 4-bromo-3-methylbenzoic acid (8.6 g, 40.0 mmol) in CH₂Cl₂ was addedDMF (0.4 mL), followed by oxalyl chloride (3.7 mL, 42 mmol) slowly over10 minutes by syringe. The reaction was stirred for 1 hour at roomtemperature, and then concentrated to give the title compound.

Step 2: 2-(4-Bromo-3-methyl-benzoyl)-3-[(E)-methylimino]-butyric acidmethyl ester

To 4-bromo-3-methyl-benzoyl chloride (40.0 mmoml) in tertahydrofuran(300 mL) wad added 3-methylamino-but-2-enoic acid methyl ester (5.12 g,40.0 mmol), followed by pyridine (3.2 mL, 40.0 mmol), and the reactionwas stirred overnight at room temperature. Solid pyridine hydrochloridecoated the flask, so the mixture was decanted, and the solid was washedthree times with EtOAc. The combined solutions were concentrated anddiluted with EtOAc (500 mL), and the mixture was washed three times withwater. The aqueous layer was separated and back-extracted with EtOAc,and the combined organic layers were dried over MgSO₄, filtered, andconcentrated to give the title compound.

Step 3: 5-(4-Bromo-3-methyl-phenyl)-3-methyl-isoxazole-4-carboxylic acidmethyl ester

2-(4-Bromo-3-methyl-benzoyl)-3-[(E)-methylimino]-butyric acid methylester (40.0 mmol) and hydroxylamine hydrochloride (2.9 g, 41 mmol) werecombined in acetic acid (100 mL) and stirred at 60° C. overnight. Themixture was concentrated, and the residue was partitioned between EtOAcand water. Saturated aqueous NaHCO₃ was added to neutralize residualacetic acid, and the organic layer was separated and washed twice withwater. The combined aqueous layers were back-extracted with EtOAc, andthe combined organic layers were dried, filtered, and concentrated. Thecrude material was purified by silica gel chromatography to give thetitle compound.

Step 4: 5-(4-Bromo-3-methyl-phenyl)-3-methyl-isoxazole-4-carboxylic acid

5-(4-Bromo-3-methyl-phenyl)-3-methyl-isoxazole-4-carboxylic acid methylester (7.26 g, 23.4 mmol) was hydrolyzed with 1N aqueous LiOH. Acidicwork-up provided the title compound.

Step 5: [5-(4-Bromo-3-methyl-phenyl)-3-methyl-isoxazol-4-yl]-carbamicacid (R)-1-(2-fluoro-phenyl)-ethyl ester

5-(4-Bromo-3-methyl-phenyl)-3-methyl-isoxazole-4-carboxylic acid (1.78g, 6.0 mmol) was dissolved in toluene (50 mL). Triethylamine (0.92 mL,6.6 mmol) was added, followed by diphenylphosphoryl azide (1.43 mL, 6.6mmol), and the mixture was stirred for 10 minutes.(R)-1-(2-Fluoro-phenyl)-ethanol (0.924 g, 6.6 mmol) was added, and thereaction was stirred at 85° C. for 3 hours, and then overnight at roomtemperature. The mixture was concentrated, and the residue was dilutedwith dichloromethane and purified by silica gel chromatography to givethe title compound.

Step 6:(4′-{4-[(R)-1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-2′-methyl-biphenyl-4-yl)-aceticacid ethyl ester

[5-(4-Bromo-3-methyl-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid(R)-1-(2-fluoro-phenyl)-ethyl ester (0.350 g, 0.80 mmol),[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic acidethyl ester (0.292 g, 0.96 mmol), and sodium bicarbonate (0.235 g, 2.8mmol) were combined in 2:1 DME:water (11 mL), and the solution waspurged with N₂ for 5 minutes. Bis(triphenylphosphine)palladium(II)dichloride (0.030 g, 0.04 mmol) was added, and the reaction was stirredat 80° C. for 2 hour. After work-up, the crude material was purified bysilica gel chromatography to give the title compound.

Step 7:(4′-{4-[(R)-1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-2′-methyl-biphenyl-4-yl)-aceticacid

(4′-{4-[(R)-1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-2′-methyl-biphenyl-4-yl)-aceticacid ethyl ester (0.282 g, 0.53 mmol) was hydrolyzed with lithiumhydroxide in methanol and water, and the crude material was purified bypreparative HPLC to give the title compound.

Example 37 Synthesis of2-(4′-{4-[1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-2-methyl-propionicacid (Compound 26) Step 1:2-(4′-{4-[1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-2-methyl-propionicacid ethyl ester

Prepared as described in Example 36, Step 6 using{3-methyl-5-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-isoxazol-4-yl}-carbamicacid 1-(2-fluoro-phenyl)-ethyl ester and2-(4-bromo-phenyl)-2-methyl-propionic acid ethyl ester; purification bypreparative HPLC was utilized.

Step 2:2-(4′-{4-[1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-2-methyl-propionicacid

2-(4′-{4-[1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-2-methyl-propionicacid ethyl ester (0.51 mmol) in Methanol (50 mL) was treated withlithium hydroxide (excess) as well as 1N aqueous LiOH (excess, and thereaction was stirred at room temperature. Analytical LCMS indicated thatstarting material was still present, so the reaction was stirred at 80°C. overnight. The mixture was neutralized with 1N HCl (20 mL) andextracted with EtOAc. The combined organic layers were washed withwater, dried, and concentrated, and the residue was purified bypreparative HPLC to give the title compound.

Example 38 Synthesis of(4′-{4-[(R)-1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid (Compound 27) Step 1: (R)-1-(2-Chloro-phenyl)-ethanol

2′-Chloroacetophenone (8.4 mL, 65 mmol) and(S)-(−)-2-methyl-CBS-oxazaborolidine (0.90 g, 0.32 mmol) were combinedin THF (75 mL). Borane methyl sulfide complex (2M in THF; 21.5 mL, 43mmol) was added over 20 minutes, and the reaction was stirred at roomtemperature for 30 minutes. MeOH was added, and the mixture wasworked-up with CH₂Cl₂ and H₂O. The organic layer was concentrated togive the title compound.

Step 2: [5-(4-Bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid(R)-1-(2-chloro-phenyl)-ethyl ester

5-(4-Bromo-phenyl)-3-methyl-isoxazole-4-carboxylic acid (10 g, 35 mmol),(R)-1-(2-chloro-phenyl)-ethanol (6.6 g, 42 mmol), triethylamine (10 mL,70 mmol), and diphenylphosphoryl azide (11.5 g, 42 mmol) were combinedin toluene (100 mL) and stirred at 90° C. for 1 hour. The mixture wasconcentrated, and the residue was purified by silica gel chromatography(0-30% EtOAc in hexanes). The isolated product was recrystallized in 5:1hexanes:acetone to give the title compound in >99% e.e (by chiral HPLC.Chiracel OD 98.4% hexanes/1.6% Ethanol. Major isomer 27.9 min minorisomer 32.7 min)

Step 3: [4-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-aceticacid ethyl ester

Ethyl 4-bromophenylacetate (9.72 g, 40 mmol), bis(pinacolato)diboron(12.2 g, 48 mmol), and potassium acetate (18.8 g, 1690 mmol) werecombined in 1,4-dioxane, and the mixture was purged with N₂ for 10minutes. 1,1′-Bis(diphenylphosphino)ferrocene)-dichloropalladium(II)(3.26 g, 4.0 mmol) was added, and the reaction was heated to 80° C. for24 hours, and at room temperature for 2 days. The mixture was dilutedwith H₂O and extracted with EtOAc. The combined organic layers werewashed with 4 times with H₂O, once with brine, and then dried, filtered,and concentrated. The residue was dissolved in CH₂Cl₂ and filteredthrough Celite to remove solids. The filtrate was concentrated, and thecrude material was purified by silica gel chromatography (0-30% EtOAc inhexanes) to give the title compound.

Step 4:(4′-{4-[(R)-1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid ethyl ester

[5-(4-Bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid(R)-1-(2-chloro-phenyl)-ethyl ester (3.0 g, 6.9 mmol),[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic acidethyl ester (2.2 g, 7.6 mmol),dichloro-bis(triphenylphosphine)palladium(II) (0.25 g, 0.35 mmol), andsodium bicarbonate (1.7 g, 20.7 mmol) were combined in DME (30 mL) andH₂O (10 mL), and the reaction was stirred at 80° C. for 2 hours. Afteraqueous work-up, the crude material was purified by silica gelchromatography (0-40% EtOAc in hexanes) to give the title compound.

Step 5:(4′-{4-[(R)-1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid

4′-{4-[(R)-1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid ethyl ester (3 g, 5.9 mmol) in MeOH and H₂O was treated withlithium hydroxide (1 g, 23.8 mmol), and the reaction was stirred at 60°C. Acidic work-up provided the title compound.

Example 39 Synthesis of2-(4′-{4-[(R)-1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-2-methyl-propionicacid (Compound 28) Step 1:2-Methyl-2-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-propionicacid ethyl ester

Prepared as described in Example 18 using2-(4-bromo-phenyl)-2-methyl-propionic acid ethyl ester andbis(pinacolato)diboron.

Step 2:2-(4′-{4-[(R)-1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-2-methyl-propionicacid ethyl ester

Prepared as described in Example 36, Step 6 using[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid(R)-1-(2-chloro-phenyl)-ethyl ester and2-methyl-2-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-propionicacid ethyl ester.

Step 3:2-(4′-{4-[(R)-1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-2-methyl-propionicacid

Prepared as described in Example 17, Step 3 using2-(4′-{4-[(R)-1-(2-chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-2-methyl-propionicacid ethyl ester.

Example 40 Synthesis of2-(4′-{4-[(R)-1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-2-methyl-propionicacid (Compound 29)

Following the procedure described in Example 36, Step 6,[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid(R)-1-(2-fluoro-phenyl)-ethyl ester and2-methyl-2-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-propionicacid ethyl ester were reacted to provide2-(4′-{4-[(R)-1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-2-methyl-propionicacid ethyl ester, which was hydrolyzed to the acid as described inExample 17, Step 3.

Example 41 Synthesis of2-(4′-{4-[(R)-1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-propionicacid (Compound 30)

2-[4-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-propionicacid ethyl ester was prepared according to the procedure described inExample 18 using 2-(4-bromo-phenyl)-propionic acid ethyl ester andbis(pinacolato)diboron.

Following the procedure described in Example 36, Step 6,[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid(R)-1-(2-fluoro-phenyl)-ethyl ester and2-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-propionicacid ethyl ester were reacted to provide2-(4′-{4-[(R)-1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-propionicacid ethyl ester, which was hydrolyzed to the acid as described inExample 17, Step 3.

Example 42 Synthesis of2-(4′-{4-[(R)-1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-propionicacid (Compound 31)

Following the procedure described in Example 36, Step 6,[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid(R)-1-(2-chloro-phenyl)-ethyl ester and2-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-propionicacid ethyl ester were reacted to provide2-(4′-{4-[(R)-1-(2-chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-propionicacid ethyl ester, which was hydrolyzed to the acid as described inExample 17, Step 3.

Example 43 Synthesis of[5-(4′-Cyanomethyl-biphenyl-4-yl)-3-methyl-isoxazol-4-yl]-carbamic acid(R)-1-(2-chloro-phenyl)-ethyl ester (Compound 61)

Prepared as described in Example 36, Step 6 using[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid(R)-1-(2-chloro-phenyl)-ethyl ester and 4-cyanomethylphenylboronic acid.

Example 44 Synthesis of[5-(4′-Cyanomethyl-biphenyl-4-yl)-3-methyl-isoxazol-4-yl]-carbamic acid(R)-1-(2-fluoro-phenyl)-ethyl ester (Compound 62)

Prepared as described in Example 36, Step 6 using[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid(R)-1-(2-fluoro-phenyl)-ethyl ester and 4-cyanomethylphenylboronic acid.

Example 45 Synthesis of{3-Methyl-5-[4′-(2H-tetrazol-5-ylmethyl)-biphenyl-4-yl]-isoxazol-4-yl}-carbamicacid (R)-1-(2-fluoro-phenyl)-ethyl ester (Compound 63)

To a solution of[5-(4′-cyanomethyl-biphenyl-4-yl)-3-methyl-isoxazol-4-yl]-carbamic acid(R)-1-(2-fluoro-phenyl)-ethyl ester (0.200 g, 0.44 mmol) in toluene (5mL) was added dibutyltin oxide (0.011 g, 0.04 mmol), followed byazidotrimethylsilane (0.07 mL, 0.53 mmol), and the reaction was stirredat 100° C. for 5 hours, and then stirred at room temperature overnight.The mixture was concentrated, and the residue was dissolved in EtOAc andwashed with water and brine. The organic layer was dried andconcentrated, and the crude material was purified by preparative HPLC togive the title compound.

Example 46 Synthesis of{3-Methyl-5-[4′-(2H-tetrazol-5-ylmethyl)-biphenyl-4-yl]-isoxazol-4-yl}-carbamicacid (R)-1-(2-chloro-phenyl)-ethyl ester (Compound 64)

Prepared as described in Example 45 using[5-(4′-cyanomethyl-biphenyl-4-yl)-3-methyl-isoxazol-4-yl]-carbamic acid(R)-1-(2-chloro-phenyl)-ethyl ester and azidotrimethylsilane.

Example 47 Synthesis of{4′-[3-Methyl-4-((R)-1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl]-biphenyl-4-yl}-aceticacid (Compound 37) Step 1: (R)-1-Phenyl-ethanol

To acetophenone (19.47 mL, 166.6 mmol) in THF (100 mL) was added(S)-(−)-2-methyl-CBS-oxazaborolidine (4.62 g, 16.6 mmol), and thereaction was cooled to 0° C. Borane methyl sulfide complex (2M in THF;50 mL, 100 mmol) was added over 15 minutes, and the reaction was stirredat room temperature. Aqueous work-up gave the title compound.

Step 2: [5-(4-Bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid(R)-1-phenyl-ethyl ester

5-(4-Bromo-phenyl)-3-methyl-isoxazole-4-carboxylic acid (25 g, 88.7mmol) in toluene (500 mL) was added triethylamine (18.5 mL, 133 mmol),followed by diphenylphosphoryl azide (22.1 mL, 101.9 mmol).(R)-(+)-1-Phenylethyl alcohol (11.9 mL, 97.5 mmol) was added, and thereaction was stirred at 75° C. for 2 hours. The mixture was partitionedbetween EtOAc and H₂O and filtered through Celite. The aqueous layer wasextracted with EtOAc, and the combined organic layers were dried overMgSO₄, filtered, and concentrated to give the title compound.

Step 3:{4′-[3-Methyl-4-((R)-1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl]-biphenyl-4-yl}-aceticacid ethyl ester

[5-(4-Bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid(R)-1-phenyl-ethyl ester (39 g, 97.2 mmol),[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic acidethyl ester (31 g, 107 mmol), and sodium bicarbonate (32.6 g, 389 mmol)were combined in 3:1 DME:H₂O (500 mL), and the mixture was purged withN₂ for 15 minutes.(1,1′-Bis(diphenylphosphino)ferrocene)-dichloropalladium(II) (2.13 g,2.91 mmol) was added, and the reaction was purged with N₂ for anadditional 10 minutes and then stirred at 90° C. overnight. The mixturewas partitioned between EtOAc and H₂O, and the aqueous layer wasextracted with EtOAc. The combined organic layers were washed with H₂O,dried over MgSO₄, filtered, and concentrated, and the residue waspurified by silica gel chromatography (EtOAc/hexane gradient) to givethe title compound.

Step 4:{4′-[3-Methyl-4-((R)-1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl]-biphenyl-4-yl}-aceticacid

To a suspension of{4′-[3-methyl-4-((R)-1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl]-biphenyl-4-yl}-aceticacid ethyl ester (24 g, 49 mmol) in 3:1 MeOH:H₂O (300 mL) was addedlithium hydroxide (8.3 g, 198 mmol), and the reaction was stirred atroom temperature overnight. The mixture was acidified and partitionedbetween EtOAc and H₂O. The aqueous layer was extracted with EtOAc, andthe combined organic layers were washed with H₂O, dried over MgSO₄,filtered, and concentrated to give the title compound. Mass spec. data(M+H)=457.

Example 48 Synthesis of{4′-[3-Methyl-4-((R)-1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl]-biphenyl-3-yl}-aceticacid (Compound 43)

(3-Bromo-phenyl)-acetic acid ethyl ester and bis(pinacolato)diboron werereacted as described in Example 18 to provide[3-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic acidethyl ester.

Following the procedure described in Example 17, Step 2,[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid(R)-1-phenyl-ethyl ester,[3-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic acidethyl ester, and bis(triphenylphosphine)palladium(II) dichloride werereacted to provide{4′-[3-methyl-4-((R)-1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl]-biphenyl-3-yl}-aceticacid ethyl ester, which was hydrolyzed to the acid as described inExample 17, Step 3.

Example 49 Synthesis of4′-[3-Methyl-4-((R)-1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl]-biphenyl-4-carboxylicacid (Compound 44)

Following the procedure described in Example 17, Step 2,[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid(R)-1-phenyl-ethyl ester and 4-ethoxycarbonylphenylboronic acid werereacted to provide4′-[3-methyl-4-((R)-1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl]-biphenyl-4-carboxylicacid ethyl ester, which was hydrolyzed to the acid as described inExample 17, Step 3.

Example 50 Synthesis of(4′-{4-[1-(2,6-Dichloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid (Compound 32)

Following the procedure described in Example 36, Step 5,5-(4′-ethoxycarbonylmethyl-biphenyl-4-yl)-3-methyl-isoxazole-4-carboxylicacid and 1-(2,6-dichlorophenyl)ethanol were reacted to provide(4′-{4-[1-(2,6-dichloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid ethyl ester, which was hydrolyzed to the acid as described inExample 34, Step 2.

Example 51 Synthesis of2-(4′-{4-[(R)-1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-2′-methyl-biphenyl-4-yl)-propionicacid (Compound 33)

Following the procedure described in Example 36, Step 6,[5-(4-bromo-3-methyl-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid(R)-1-(2-fluoro-phenyl)-ethyl ester and2-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-propionicacid ethyl ester were reacted to provide2-(4′-{4-[(R)-1-(2-fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-2′-methyl-biphenyl-4-yl)-propionicacid ethyl ester, which was hydrolyzed to the acid as described inExample 36, Step 7.

Example 52 Synthesis of(4′-{4-[(S)-1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid (Compound 34)

(S)-1-(2-Fluoro-phenyl)-ethanol was prepared according to the proceduredescribed in Example 35, Step 1 using 2′-fluoroacetophenone and(R)-(+)-2-methyl-CBS-oxazaborolidine.

Following the procedure described in Example 36, Step 5,5-(4′-ethoxycarbonylmethyl-biphenyl-4-yl)-3-methyl-isoxazole-4-carboxylicacid and (S)-1-(2-fluoro-phenyl)-ethanol were reacted to provide(4′-{4-[(S)-1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid ethyl ester, which was hydrolyzed to the acid as described inExample 34, Step 2.

Example 53 Synthesis of(4′-{4-[(S)-1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid (Compound 35)

(S)-1-(2-Chloro-phenyl)-ethanol was prepared as described in Example 35,Step 1 using 2′-chloroacetophenone and(R)-(+)-2-methyl-CBS-oxazaborolidine.

[5-(4-Bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid(S)-1-(2-chloro-phenyl)-ethyl ester was prepared as described in Example36, Step 5 using5-(4′-ethoxycarbonylmethyl-biphenyl-4-yl)-3-methyl-isoxazole-4-carboxylicacid and (S)-1-(2-chloro-phenyl)-ethanol.

Following the procedure described in Example 36, Step 6,[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid(S)-1-(2-chloro-phenyl)-ethyl ester and[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic acidethyl ester were reacted to provide(4′-{-4-[(S)-1-(2-chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid ethyl ester, which was hydrolyzed to the acid as described inExample 34, Step 2.

Example 54 Synthesis of{4′-[4-(2-Chloro-benzyloxycarbonylamino)-3-methyl-isoxazol-5-yl]-biphenyl-4-yl}-aceticacid (Compound 36)

[5-(4-Bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid2-chloro-benzyl ester was prepared as described in Example 36, Step 5using 5-(4-bromo-phenyl)-3-methyl-isoxazole-4-carboxylic acid andchlorobenzyl alcohol.

Following the procedure described in Example 36, Step 6,[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid2-chloro-benzyl ester and[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic acidethyl ester were reacted to provide{4′-[4-(2-chloro-benzyloxycarbonylamino)-3-methyl-isoxazol-5-yl]-biphenyl-4-yl}-aceticacid ethyl ester, which was hydrolyzed to the acid as described inExample 36, Step 7.

Example 55 Synthesis of(4′-{4-[1-(2,3-Difluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid (Compound 38)

2′,3′-Difluoroacetophenone (0.337 g, 2.16 mmol) in methanol (15 mL) wastreated with sodium borohydride (0.090 g, 2.37 mmol), and the reactionwas stirred at room temperature for 30 minutes. The mixture waspartitioned between dichloromethane and water, and the organic layer wasseparated, dried over MgSO₄, filtered, and concentrated to give1-(2,3-difluoro-phenyl)-ethanol.

5-(4-bromo-phenyl)-3-methyl-isoxazole-4-carboxylic acid and1-(2,3-difluoro-phenyl)-ethanol were reacted as described in Example 36,Step 5 to provide [5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamicacid 1-(2,3-difluoro-phenyl)-ethyl ester.

Following the procedure described in Example 36, Step 6,[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid1-(2,3-difluoro-phenyl)-ethyl ester and[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic acidethyl ester were reacted to provide(4′-{4-[1-(2,3-difluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid ethyl ester, which was hydrolyzed to the acid as described inExample 34, Step 2.

Example 56 Synthesis of(4′-{4-[1-(2,4-Difluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid (Compound 39)

2′,4′-difluoroacetophenone was reduced to 1-(2,4-Difluorophenyl)ethanolas described in Example 55.

5-(4-bromo-phenyl)-3-methyl-isoxazole-4-carboxylic acid and1-(2,4-difluorophenyl)ethanol were reacted as described in Example 36,Step 5 to provide [5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamicacid 1-(2,4-difluoro-phenyl)-ethyl ester.

Following the procedure described in Example 36, Step 6,[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid1-(2,4-difluoro-phenyl)-ethyl ester and[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic acidethyl ester were reacted to provide(4′-{4-[1-(2,4-Difluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid ethyl ester, which was hydrolyzed to the acid as described inExample 34, Step 2.

Example 57 Synthesis of(4′-{4-[1-(2-Fluoro-4-methoxy-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid (Compound 40)

2′-Fluoro-4′-methoxyacetophenone was reduced to1-(2-fluoro-4-methoxyphenyl)ethan-1-ol as described in Example 55.

5-(4-Bromo-phenyl)-3-methyl-isoxazole-4-carboxylic acid and1-(2-fluoro-4-methoxyphenyl)ethan-1-ol were reacted as described inExample 36, Step 5 to provide[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid1-(2-fluoro-4-methoxy-phenyl)-ethyl ester.

Following the procedure described in Example 36, Step 6,[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid1-(2-fluoro-4-methoxy-phenyl)-ethyl ester and[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic acidethyl ester were reacted to provide(4′-{4-[1-(2-fluoro-4-methoxy-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid ethyl ester, which was hydrolyzed to the acid as described inExample 34, Step 2.

Example 58 Synthesis of(4′-{4-[1-(2,5-Difluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid (Compound 41)

2′,5′-Difluoroacetophenone was reduced to 1-(2,5-Difluorophenyl)ethanolas described in Example 55.

5-(4-bromo-phenyl)-3-methyl-isoxazole-4-carboxylic acid and1-(2,5-difluorophenyl)ethanol were reacted as described in Example 36,Step 5 to provide [5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamicacid 1-(2,5-difluoro-phenyl)-ethyl ester.

Following the procedure described in Example 36, Step 6,[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid1-(2,5-difluoro-phenyl)-ethyl ester and[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic acidethyl ester were reacted to provide(4′-{4-[1-(2,5-difluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid ethyl ester, which was hydrolyzed to the acid as described inExample 34, Step 2.

Example 59 Synthesis of(4′-{4-[1-(2,6-Difluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid (Compound 42)

2′,6′-difluoroacetophenone was reduced to1-(2,6-difluorophenyl)ethan-1-ol as described in Example 55.

5-(4-bromo-phenyl)-3-methyl-isoxazole-4-carboxylic acid and1-(2,6-difluorophenyl)ethan-1-ol were reacted as described in Example36, Step 5 to provide[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid1-(2,6-difluoro-phenyl)-ethyl ester.

Following the procedure described in Example 36, Step 6,[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid1-(2,6-difluoro-phenyl)-ethyl ester and[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic acidethyl ester were reacted to provide(4′-{4-[1-(2,6-difluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid ethyl ester, which was hydrolyzed to the acid as described inExample 34, Step 2.

Example 60 Synthesis of{4′-[3-Methyl-4-((R)-1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl]-biphenyl-2-yl}-aceticacid (Compound 45)

Following the procedure described in Example 36, Step 6,[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid(R)-1-phenyl-ethyl ester and[2-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic acidethyl ester were reacted to provide{4′-[3-Methyl-4-((R)-1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl]-biphenyl-2-yl}-aceticac id ethyl ester, which was hydrolyzed to the acid as described inExample 34, Step 2.

Example 61 Synthesis of{4′-[3-Methyl-4-((R)-1-o-tolyl-ethoxycarbonylamino)-isoxazol-5-yl]-biphenyl-4-yl}-aceticacid (Compound 46)

2′-methylacetophenone and (S)-(−)-2-methyl-CBS-oxazaborolidine werereacted as described in Example 35, Step 1 to provide(R)-1-o-Tolyl-ethanol.

5-(4-Bromo-phenyl)-3-methyl-isoxazole-4-carboxylic acid and(R)-1-o-tolyl-ethanol were reacted as described in Example 36, Step 5 toprovide [5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid(R)-1-o-tolyl-ethyl ester.

Following the procedure described in Example 36, Step 6,[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid(R)-1-o-tolyl-ethyl ester and[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic acidethyl ester were reacted to provide{4′-[3-methyl-4-((R)-1-o-tolyl-ethoxycarbonylamino)-isoxazol-5-yl]-biphenyl-4-yl}-aceticacid ethyl ester, which was hydrolyzed to the acid as described inExample 34, Step 2.

Example 62 Synthesis of2-(4′-{4-[(R)-1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-propionicacid (Diastereomers—Compound 47 and Compound 48) Step 1:2-(4-Bromo-phenyl)-propionic acid ethyl ester

2-(4-Bromo-phenyl)-propionic acid ethyl ester (10.0 g, 41 mmol) wasdissolved in tetrahydrofuran (400 mL) and cooled to −78° C. Sodiumbis(trimethylsilyl)amide (2M in tetrahydrofuran; 53.5 mL, 107 mmol) wasadded, and the mixture was stirred for 1 hour at −78° C. Iodomethane(2.814 mL, 45.1 mmol) was added, and the reaction was slowly warmed toroom temperature and stirred overnight. The mixture was concentrated,and the residue was extracted with EtOAc. The combined organic layerswere washed with water and brine, and then dried and concentrated. Thecrude material was purified by silica gel chromatography (0-15% EtOAc inhexanes) to give the title compound.

Step 2: 2-(4-Bromo-phenyl)-propionic acid

Prepared according to the procedure described in Example 6, Step 4 using2-(4-bromo-phenyl)-propionic acid ethyl ester.

Step 3: 2-(4-Bromo-phenyl)-propionyl chloride

2-(4-Bromo-phenyl)-propionic acid (0.500 g, 2.18 mmol) indichloromethane (20 mL) was treated with dimethylformamide (0.03 mL,0.44 mmol), followed by oxalyl chloride (0.248 mL, 2.8 mmol) dropwiseover 5 minutes, and the reaction was stirred at room temperature for 1hour. The mixture was then concentrated and dried under vacuum to givethe title compound.

Step 4:(4R,5S)-3-[2-(4-Bromo-phenyl)-propionyl]-4-methyl-5-phenyl-oxazolidin-2-one

(4R,5S)-(+)-4-Methyl-5-phenyl-2-oxazolidinone (0.348 g, 1.96 mmol) wasdissolved in tetrahydrofuran (20 mL) and cooled to −78° C.n-Butyllithium (1.6M in hexanes; 1.64 mL, 2.62 mmol) was added, and thereaction was stirred for 1 hour at −78° C. 2-(4-Bromo-phenyl)-propionylchloride (0.540 g, 2.18 mmol) in tetrahydrofuran (10 mL) was added, andthe reaction was stirred for 30 minutes. The mixture was concentrated,and the residue was partitioned between EtOAc and water. The organiclayer was separated, dried, and concentrated, and the crude material waspurified by silica gel chromatography (0-25% EtOAc in hexanes) to givetwo diastereomers as separated products. Diastereomer A was the firstproduct to come off the column, while Diastereomer B was the secondproduct to come off the column.

Step 5:(4R,5S)-4-Methyl-5-phenyl-3-{2-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-propionyl}-oxazolidin-2-one

Diastereomer A—Prepared as described in Example 5, Step 2 using(4R,5S)-3-[2-(4-bromo-phenyl)-propionyl]-4-methyl-5-phenyl-oxazolidin-2-one(Diastereomer A) and bis(pinacolato)diboron.

Diastereomer B—Prepared as described in Example 5, Step 2 using(4R,5S)-3-[2-(4-bromo-phenyl)-propionyl]-4-methyl-5-phenyl-oxazolidin-2-one(Diastereomer B) and bis(pinacolato)diboron.

Step 6:(3-Methyl-5-{4′-[1-methyl-2-((4R,5S)-4-methyl-2-oxo-5-phenyl-oxazolidin-3-yl)-2-oxo-ethyl]-biphenyl-4-yl}-isoxazol-4-yl)-carbamicacid (R)-1-(2-chloro-phenyl)-ethyl ester

Diastereomer A—Prepared as described in Example 36, Step 6 using(4R,5S)-4-methyl-5-phenyl-3-{2-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-propionyl}-oxazolidin-2-one(Diastereomer A) and[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid(R)-1-(2-chloro-phenyl)-ethyl ester.

Diastereomer B—Prepared as described in Example 36, Step 6 using(4R,5S)-4-methyl-5-phenyl-3-{2-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-propionyl}-oxazolidin-2-one(Diastereomer B) and[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid(R)-1-(2-chloro-phenyl)-ethyl ester.

Step 7:2-(4′-{4-[(R)-1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-propionicacid

Diastereomer A—Prepared as described in Example 34, Step 2 using(3-methyl-5-{4′-[1-methyl-2-((4R,5S)-4-methyl-2-oxo-5-phenyl-oxazolidin-3-yl)-2-oxo-ethyl]-biphenyl-4-yl}-isoxazol-4-yl)-carbamicacid (R)-1-(2-chloro-phenyl)-ethyl ester (Diastereomer A).

Diastereomer B—Prepared as described in Example 34, Step 2 using(3-methyl-5-{4′-[1-methyl-2-((4R,5S)-4-methyl-2-oxo-5-phenyl-oxazolidin-3-yl)-2-oxo-ethyl]-biphenyl-4-yl}-isoxazol-4-yl)-carbamicacid (R)-1-(2-chloro-phenyl)-ethyl ester (Diastereomer B)

Example 64 Synthesis of(3′-Chloro-4′-{4-[(R)-1-(2-chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid (Compound 49)

Following the procedures described in Example 36:4-bromo-2-chlorobenzoic acid and oxalyl chloride were reacted to provide4-bromo-2-chloro-benzoyl chloride, which was reacted with3-methylamino-but-2-enoic acid methyl ester to provide2-(4-bromo-2-chloro-benzoyl)-3-[(E)-methylimino]-butyric acid methylester. 2-(4-Bromo-2-chloro-benzoyl)-3-[(E)-methylimino]-butyric acidmethyl ester and hydroxylamine hydrochloride were then reacted toprovide 5-(4-bromo-2-chloro-phenyl)-3-methyl-isoxazole-4-carboxylic acidmethyl ester. Following Example 36, Step 4,5-(4-bromo-2-chloro-phenyl)-3-methyl-isoxazole-4-carboxylic acid methylester was hydrolyzed to5-(4-bromo-2-chloro-phenyl)-3-methyl-isoxazole-4-carboxylic acid.5-(4-Bromo-2-chloro-phenyl)-3-methyl-isoxazole-4-carboxylic acid and(R)-1-(2-chloro-phenyl)-ethanol were then reacted as described inExample 36, Step 5 to provide[5-(4-bromo-2-chloro-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid(R)-1-(2-chloro-phenyl)-ethyl ester.

Following the procedure described in Example 36, Step 6,[5-(4-bromo-2-chloro-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid(R)-1-(2-chloro-phenyl)-ethyl ester and[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic acidethyl ester were reacted to provide(3′-chloro-4′-{4-[(R)-1-(2-chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid ethyl ester, which was hydrolyzed to the acid as described inExample 36, Step 7.

Example 65 Synthesis of2-(4′-{4-[(R)-1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-butyricacid (Compound 50)

Prepared as described in Example 36, Step 6 using{3-methyl-5-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-isoxazol-4-yl}-carbamicacid (R)-1-(2-chloro-phenyl)-ethyl ester and 2-(4-bromophenyl)butanoicacid.

Example 66 Synthesis of(T-Chloro-4′-{4-[(R)-1-(2-chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid (Compound 51)

Following the procedures described in Example 36:4-bromo-3-chlorobenzoic acid and oxalyl chloride were reacted to provide4-bromo-3-chloro-benzoyl chloride, which was then reacted with3-methylamino-but-2-enoic acid methyl ester to provide2-(4-bromo-3-chloro-benzoyl)-3-[(E)-methylimino]-butyric acid methylester. 2-(4-Bromo-3-chloro-benzoyl)-3-[(E)-methylimino]-butyric acidmethyl ester and hydroxylamine hydrochloride were then reacted asdescribed in Example 36, Step 3 to provide5-(4-bromo-3-chloro-phenyl)-3-methyl-isoxazole-4-carboxylic acid methylester, which was then hydrolyzed to5-(4-bromo-3-chloro-phenyl)-3-methyl-isoxazole-4-carboxylic acid asdescribed in Example 36, Step 4.5-(4-Bromo-3-chloro-phenyl)-3-methyl-isoxazole-4-carboxylic acid and(R)-1-(2-chloro-phenyl)-ethanol were reacted to provide[5-(4-bromo-3-chloro-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid(R)-1-(2-chloro-phenyl)-ethyl ester as described in Example 36, Step 5.

Following the procedure described in Example 36, Step 6,[5-(4-bromo-3-chloro-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid(R)-1-(2-chloro-phenyl)-ethyl ester and[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic acidethyl ester were reacted to provide (2′-chloro-4′-{4-[(R)-1-(2-chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid ethyl ester, which was hydrolyzed to the acid as described inExample 36, Step 7.

Example 67 Synthesis of(4′-{4-[(R)-1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-2′-fluoro-biphenyl-4-yl)-aceticacid (Compound 52)

Following the procedures described in Example 36:4-bromo-3-fluorobenzoyl chloride and oxalyl chloride were reacted toprovide 4-bromo-3-fluoro-benzoyl chloride, which was then reacted with3-methylamino-but-2-enoic acid methyl ester to provide2-(4-bromo-3-fluoro-benzoyl)-3-[(E)-methylimino]-butyric acid methylester. 2-(4-bromo-3-fluoro-benzoyl)-3-[(E)-methylimino]-butyric acidmethyl ester and hydroxylamine hydrochloride were then reacted asdescribed in Example 36, Step 3 to provide5-(4-bromo-3-fluoro-phenyl)-3-methyl-isoxazole-4-carboxylic acid methylester, which was then hydrolyzed to5-(4-bromo-3-fluoro-phenyl)-3-methyl-isoxazole-4-carboxylic acid asdescribed in Example 36, Step 4.5-(4-Bromo-3-fluoro-phenyl)-3-methyl-isoxazole-4-carboxylic acid and(R)-1-(2-chloro-phenyl)-ethanol were reacted to provide[5-(4-Bromo-3-fluoro-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid(R)-1-(2-chloro-phenyl)-ethyl ester as described in Example 36, Step 5.

Following the procedure described in Example 36, Step 6,[5-(4-bromo-3-fluoro-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid(R)-1-(2-chloro-phenyl)-ethyl ester and[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic acidethyl ester were reacted to provide(4′-{4-[(R)-1-(2-chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-2′-fluoro-biphenyl-4-yl)-aceticacid ethyl ester, which was hydrolyzed to the acid as described inExample 36, Step 7.

Example 68 Synthesis of4′-{4-[(R)-1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-carboxylicacid (Compound 53)

Following the procedure described in Example 36, Step 6,[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid(R)-1-(2-chloro-phenyl)-ethyl ester and 4-ethoxycarbonylphenylboronicacid were reacted to provide4′-{4-[(R)-1-(2-chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-carboxylicacid ethyl ester, which was hydrolyzed to the acid as described inExample 36, Step 7.

Example 71 Synthesis of(4′-{4-[(R)-1-(3,5-Dibromo-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid (Compound 56)

To a solution of 3,5-dibromobenzoic acid (2.5 g, 8.9 mmol) in Et₂O (30mL) at 0° C. was added methyl lithium (1.6M in diethyl ether; 12.3 mL,19.6 mmol) was added dropwise. The reaction was warmed to roomtemperature and stirred for 2 hours. Acidic work-up, followed by silicagel chromatography, gave 1-(3,5-dibromo-phenyl)-ethanone.

1-(3,5-dibromo-phenyl)-ethanone and (S)-(−)-2-methyl-CBS-oxazaborolidinewere reacted as described in Example 35, Step 1 to provide(R)-1-(3,5-dibromo-phenyl)-ethanol.

Following the procedure described in Example 36, Step 5,5-(4′-ethoxycarbonylmethyl-biphenyl-4-yl)-3-methyl-isoxazole-4-carboxylicacid and (R)-1-(3,5-dibromo-phenyl)-ethanol were reacted to provide(4′-{4-[(R)-1-(3,5-dibromo-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid ethyl ester, which was hydrolyzed to the acid as described inExample 34, Step 2.

Example 72 Synthesis of{4′-[3-Methyl-4-((S)-1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl]-biphenyl-4-yl}-aceticacid (Compound 57)

2′-Chloroacetophenone and (R)-(+)-2-methyl-CBS-oxazaborolidine werereacted as described in Example 35, Step 1 to provide(S)-1-phenyl-ethanol.

5-(4-bromo-phenyl)-3-methyl-isoxazole-4-carboxylic acid and(S)-1-phenyl-ethanol were reacted as described in Example 36, Step 5 toprovide [5-(4-Bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid(S)-1-phenyl-ethyl ester.

Following the procedure described in Example 36, Step 6,[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid(S)-1-phenyl-ethyl ester and[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic acidethyl ester were reacted to provide{4′-[3-methyl-4-((S)-1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl]-biphenyl-4-yl}-aceticacid ethyl ester, which was hydrolyzed to the acid as described inExample 34, Step 2.

Example 73 Synthesis of(4′-{4-[(R)-1-(3-Hydroxy-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid (Compound 58) 3′-hydroxyacetophenone and(S)-(−)-2-methyl-CBS-oxazaborolidine were reacted as described inExample 35, Step 1 to provide 3-((R)-1-hydroxy-ethyl)-phenol

5-(4-bromo-phenyl)-3-methyl-isoxazole-4-carboxylic acid and3-((R)-1-hydroxy-ethyl)-phenol were reacted as described in Example 36,Step 5 to provide [5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamicacid (R)-1-(3-hydroxy-phenyl)-ethyl ester.

Following the procedure described in Example 36, Step 6,[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid(R)-1-(3-hydroxy-phenyl)-ethyl ester and[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic acidethyl ester were reacted to provide(4′-{4-[(R)-1-(3-hydroxy-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid ethyl ester, which was hydrolyzed to the acid as described inExample 34, Step 2.

Example 74 Synthesis of{4′-[3-Methyl-4-(1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl]-biphenyl-4-yl}-aceticacid (Compound 59)

5-(4-Bromo-phenyl)-3-methyl-isoxazole-4-carboxylic acid and1-phenylethanol were reacted as described in Example 36, Step 5 toprovide [5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid1-phenyl-ethyl ester.

Following the procedure described in Example 36, Step 6,[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid 1-phenyl-ethylester and[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic acidethyl ester were reacted to provide{4′-[3-methyl-4-(1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl]-biphenyl-4-yl}-aceticacid ethyl ester, which was hydrolyzed to the acid as described inExample 34, Step 2.

Example 75 Synthesis of{4′-[3-Methyl-4-(1-phenyl-ethoxy-d9-carbonylamino)-isoxazol-5-yl]-biphenyl-4-yl}-aceticacid (Compound 60)

5-(4-bromo-phenyl)-3-methyl-isoxazole-4-carboxylic acid and1-phenylethanol-d9 (fully dueterated 1-phenylethanol obtained fromCarbocore) were reacted as described in Example 36, Step 5 to provide[5-(4-Bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid1-phenyl-ethyl-d9 ester.

Following the procedure described in Example 36, Step 6,[5-(4-bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid1-phenyl-ethyl-d9 ester and[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-acetic acidethyl were reacted to provide{4′-[3-methyl-4-(1-phenyl-ethoxy-d9-carbonylamino)-isoxazol-5-yl]-biphenyl-4-yl}-aceticacid ethyl ester, which was hydrolyzed to the acid as described inExample 34, Step 2.

Example 76 Synthesis of[5-(4′-Carbamimidoylmethyl-biphenyl-4-yl)-3-methyl-isoxazol-4-yl]-carbamicacid (R)-1-(2-fluoro-phenyl)-ethyl ester (Compound 65)

Step 1: [5-(4-Bromo-phenyl)-3-methyl-isoxazol-4-yl]-carbamic acid(R)-1-(2-fluoro-phenyl)-ethyl ester and 4-cyanomethylphenylboronic acidwere reacted as described in Example 36, Step 6 to provide[5-(4′-cyanomethyl-biphenyl-4-yl)-3-methyl-isoxazol-4-yl]-carbamic acid(R)-1-(2-fluoro-phenyl)-ethyl ester.

Step 2:[5-(4′-Cyanomethyl-biphenyl-4-yl)-3-methyl-isoxazol-4-yl]-carbamic acid(R)-1-(2-fluoro-phenyl)-ethyl ester (0.400 g, 0.88 mmol) in ethanol (5mL) was treated with 4N HCl in 1,4-dioxane (5 mL), and the reaction wasstirred overnight at room temperature. The mixture was concentrated todryness, and then dissolved in 2M NH₃ in methanol. The reaction wasstirred overnight at room temperature, and then additional 2M NH₃ inmethanol was added and the reaction was stirred at 40° C. for 1.5 hours.The mixture was concentrated, and the residue was purified bypreparative HPLC to give the title compound.

Example 77 Synthesis of{5-[4′-(2-Acetylamino-2-imino-ethyl)-biphenyl-4-yl]-3-methyl-isoxazol-4-yl}-carbamicacid (R)-1-(2-fluoro-phenyl)-ethyl ester (Compound 66)

To[5-(4′-carbamimidoylmethyl-biphenyl-4-yl)-3-methyl-isoxazol-4-yl]-carbamicacid (R)-1-(2-fluoro-phenyl)-ethyl ester (0.055 g, 0.12 mmol) indichloromethane (5 mL) was added diisopropylethylamine (0.052 mL, 0.3mmol), followed by acetyl chloride (0.009 mL, 0.126 mmol), and thereaction was stirred at room temperature. Additional acetyl chloride(0.009 mL, 0.126 mmol) was added, and the reaction was stirred at roomtemperature. The mixture was quenched with water and extracted withdichloromethane. The combined organic layers were dried, filtered, andconcentrated, and the residue was purified by preparative HPLC to givethe title compound.

Example 78 Synthesis of[4′-(4-Amino-3-methyl-isoxazol-5-yl)-biphenyl-4-yl]acetic acid

[4′-(4-tert-Butoxycarbonylamino-3-methyl-isoxazol-5-yl)-biphenyl-4-yl]-aceticacid (1.0 mmol) was treated with trifluoroacetic acid (5 mL) for 1 hour.Work-up provided the title compound. Mass spec. data (M+H)=309.

Example 79 Synthesis of2-(2-{4′-[3-Methyl-4-((R)-1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl]-biphenyl-4-yl}-acetylamino)-ethanesulfonicacid (Compound 67)

Rats were dosed with{4′-[3-ethyl-4-((R)-1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl]-biphenyl-4-yl}-aceticacid (30 mg/kg) and the bile was collected over 6 hours. The titlecompound was purified from bile by reverse phase HPLC. Mass spec. data(M+H)=564.

In some embodiments, Mass spectrometric data (mass spec. data) isobtained on with a Shimadzu LCMS 2010A.

In vitro Assay(s)

Example 80 Establishment of a CHO Cell Line Stably Expressing Human LPA₁

A 1.1 kb cDNA encoding the human LPA₁ receptor was cloned from humanlung. Human lung RNA (Clontech Laboratories, Inc. USA) was reversetranscribed using the RETROscript kit (Ambion, Inc.) and the full-lengthcDNA for human LPA₁ was obtained by PCR of the reverse transcriptionreaction. The nucleotide sequence of the cloned human LPA₁ wasdetermined by sequencing and confirmed to be identical to the publishedhuman LPA₁ sequence (An et al. Biochem. Biophys. Res. Commun. 231:619(1997). The cDNA was cloned into the pcDNA5/FRT expression plasmid andtransfected in CHO cells using lipofectamine 2000 (Invitrogen Corp.,USA). Clones stably expressing human LPA₁ were selected using hygromycinand identified as cells that show Ca-influx in response to LPA.

Example 81 Generation of Cells Transiently Expressing Human LPA₂

A vector containing the human LPA₂ receptor cDNA was obtained from theMissouri S&T cDNA Resource Center (www.cdna.org). The full-length cDNAfragment for human LPA₂ was obtained by PCR from the vector. Thenucleotide sequence of the cloned human LPA₂ was determined bysequencing and confirmed to be identical to the published human LPA₂sequence (NCBI accession number NM_(—)004720). The cDNA was cloned intothe pcDNA3.1 expression plasmid and transfected into B103 cells(Invitrogen Corp., USA) by seeding cells in a 96-well poly-D-lysinecoated plate at 30,000-35,000 cells per well together with 0.2 μllipofectamine 2000 and 0.2 lag of the LPA₂ expression vector. Cells werecultured overnight in complete media before being assayed forLPA-induced Ca-influx.

Example 82 Establishment of a CHO Cell Line Stably Expressing Human LPA₃

A vector containing the human LPA₃ receptor cDNA was obtained from theMissouri S&T cDNA Resource Center (www.cdna.org). The full-length cDNAfragment for human LPA₃ was obtained by PCR from the vector. Thenucleotide sequence of the cloned human LPA₃ was determined bysequencing and confirmed to be identical to the published human LPA₃sequence (NCBI accession number NM_(—)012152). The cDNA was cloned intothe pcDNA5/FRT expression plasmid and transfected in CHO cells usinglipofectamine 2000 (Invitrogen Corp., USA). Clones stably expressinghuman LPA₃ were selected using hygromycin and identified as cells thatshow Ca-influx in response to LPA.

Example 83 LPA1 and LPA3 Calcium Flux Assays

Human LPA₁ or LPA₃ expressing CHO cells are seeded at 20,000-45,000cells per well in a 96-well poly-D-lysine coated plate one or two daysbefore the assay. Prior to the assay, the cells are washed once with PBSand then cultured in serum-free media overnight. On the day of theassay, a calcium indicator dye (Calcium 4, Molecular Devices) in assaybuffer (HBSS with Ca²⁺ and Mg²⁺ and containing 20 mM Hepes and 0.3%fatty-acid free human serum albumin) is added to each well andincubation continued for 1 hour at 37° C. 10 μl of test compounds in2.5% DMSO are added to the cells and incubation continued at roomtemperature for 30 minutes. Cells are the stimulated by the addition of10 nM LPA and intracellular Ca²⁺ measured using the Flexstation 3(Molecular Devices). IC₅₀s are determined using Graphpad prism analysisof drug titration curves.

Example 84 LPA2 Calcium Flux Assay

Following an overnight culture with lipofectamine 2000 and the LPA₂expression vector, the B103 cells are washed once with PBS then serumstarved for 4 hours. A calcium indicator dye (Calcium 4, MolecularDevices) in assay buffer (HBSS with Ca²⁺ and Mg²⁺ and containing 20 mMHepes and 0.3% fatty-acid free human serum albumin) is added to eachwell and incubation continued for 1 hour at 37° C. 10 μl of testcompounds in 2.5% DMSO are added to the cells and incubation continuedat room temperature for 30 minutes. Cells are the stimulated by theaddition of 10 nM LPA and intracellular Ca²⁺ measured using theFlexstation 3 (Molecular Devices). IC₅₀s are determined using Graphpadprism analysis of drug titration curves.

Example 85 GTPγS Binding Assay

The ability of a compound to inhibit binding of GTP to LPA₁ is assessedvia a membrane GTPγS assay. CHO cells stably expressing the recombinanthuman LPA₁ receptor are resuspended in 10 mM Hepes, 7.4 containing 1 mMDTT, lysed and centrifuged at 75,000×g to pellet the membranes. Themembranes are resuspended in 10 mM Hepes, 7.4 containing 1 mM DTT and10% glycerol. Membranes (−25 μg per well) are incubated in 96-wellplates with 0.1 nM [³⁵S]-GTPγS, 900 nM LPA, 5 μM GDP, and test compoundin Assay Buffer (50 mM Hepes, pH 7.4, 100 mM NaCl, 10 mM MgCl₂, 50 μg/mlsaponin and 0.2% fatty-acid free human serum albumin) for 30 minutes at30° C. The reactions are terminated by rapid filtration through WhatmanGF/B glass fibre filter plates. The filter plates are washed 3 timeswith 1 ml cold Wash Buffer (50 mM Hepes, 7.5, 100 mM NaCl and 10 mMMgCl₂) and dried. Scintillant is then added to the plates and theradioactivity retained on the filters is determined on a PackardTopCount (Perkin Elmer). Specific binding is determined as totalradioactive binding minus non-specific binding in the absence of theligand (900 nM LPA). IC₅₀s were determined using Graphpad prism analysisof drug titration curves.

Illustrative in vitro biological data for representative compounds ofFormula (I) is presented in the Table below. Unless otherwise noted,compounds that were tested had an IC₅₀ of less than 50 μM in the HLPA1Ca Flux assay.

Compound HLPA1 Ca HLPA3 Ca Flux Number Flux IC50 (uM) IC50 (uM) 1 A C 2B C 3 A C 4 A C 5 A C 6 A C 7 C C 8 A C 9 C ND 10 A C 11 A C 12 A B 13 AC 14 A C 15 A C 16 A C 17 A ND 18 A C 19 A ND 20 A C 21 A ND 22 A C 23 AC 24 A C 25 A C 26 A 27 A C 28 A B 29 A C 30 A C 31 A B 32 C ND 33 A ND34 A C 35 B C 36 B ND 37 A C 38 A C 39 A C 40 C ND 41 A C 42 A D 43 B D44 A C 45 C ND 46 A C 47 A B 48 A C 49 A C 50 A B 51 A C 52 A C 53 A B56 C ND 57 A C 58 C D 61 A C 62 A D 63 A C 64 A C 65 C ND 66 A ND 67 AND A = less than 0.3 μM; B = greater than 0.3 μM and less than 1 μM; C =greater than 1 μM and less than 50 μM; D = greater than 50 μM; ND = notdetermined.

Example 86 LPA1 Chemotaxis Assay

Chemotaxis of the A2058 human melanoma cells was measured using theNeuroprobe ChemoTx® System plates (8 μm pore size, 5.7 mm diametersites). The filter sites were coated with 0.001% fibronectin (Sigma) in20 mM Hepes, pH 7.4 and allowed to dry. A2058 cells were serum-starvedfor 24 hours, then harvested with Cell Stripper and resuspended in DMEMcontaining 0.1% fatty-acid-free bovine serum albumin (BSA) to aconcentration of 1×10⁶/ml. Cells were mixed with an equal volume of testcompound (2×) in DMEM containing 0.1% fatty-acid-free BSA and incubatedat 37° C. for 15 minutes. LPA (100 nM in DMEM containing 0.1%fatty-acid-free BSA) or vehicle was added to each well of the lowerchamber and 50 μl of the cell suspension/test compound mix was appliedto the upper portion of the ChemoTx plate. Plates were incubated at 37°C. for three hours and then the cells removed from the upper portion byrinsing with PBS and scraping. The filter was dried then stained withHEMA 3 Staining System (Fisher Scientific). The absorbance of the filterwas read at 590 nM and IC₅₀s were determined using Symyx Assay Explorer.

Compound 27 and Compound 37 inhibited LPA-driven chemotaxis (IC₅₀ lessthan 300 nM) of human A2058 melanoma cells.

In Vivo Assay(s)

Example 87 Bleomycin-Induced Lung Fibrosis Model in Mice

Female CD-1 mice (Harlan, 25-30 g) are housed 4 per cage, given freeaccess to food and water and allowed to acclimate for at least 7 daysprior to test initiation. After the habituation phase, mice are lightlyanesthetized with isoflurane (5% in 100% O₂) and administered withbleomycin sulfate (0.01-5 U/kg, Henry Schein) via intratrachealinstillation (Cuzzocrea S et al. Am J Physiol Lung Cell Mol. Physiol.2007 May; 292(5):L1095-104. Epub 2007 Jan. 12). Mice are returned totheir cages and monitored daily for the duration of the experiment. Testcompound or vehicle is delivered po, ip or sc daily. The route andfrequency of dosing is based on previously determined pharmacokineticproperties. All animals are sacrificed using inhaled isoflurane 3, 7,14, 21 or 28 days after bleomycin instillation. Following sacrifice,mice are intubated with a 20 gauge angiocatheter attached to a 1 mlsyringe. Lungs are lavaged with saline to obtain bronchoalveolar lavagefluid (BALF) and then removed and fixed in 10% neutral buffered formalinfor subsequent histopathological analysis. BALF is centrifuged for 10min at 800×g to pellet the cells and the cell supernatant removed andfrozen at −80° C. for subsequent protein analysis using the DC proteinassay kit (Biorad, Hercules, Calif.) and soluble collagen analysis usingSircol (Biocolor Ltd, UK). BALF is analyzed for concentrations ofinflammatory, pro-fibrotic and tissue injury biomarkers includingtransforming growth factor β1, hyaluronic acid, tissue inhibitor ofmetalloproteinase-1, matrix matelloproteinase-7, connective tissuegrowth factor and lactate dehydrogenase activity, using commerciallyavailable ELISA. The cell pellet is re-suspended in PBS. Total cellcounts are then obtained using a Hemavet hematology system (DrewScientific, Wayne, Pa.) and differential cells counts are determinedusing Shandon cytospin (Thermo Scientific, Waltham, Mass.). Lung tissueis stained using hematoxylin and eosin (H&E) and trichrome and lungfibrosis is determined by semiquantitative histopathological scoring(Ashcroft T. et al. J. Clin. Path. 1988; 41; 4, 467-470) using lightmicroscopy (10× magnification) and quantitative, computer-assisteddensitometry of collagen in lung tissue sections using light microscopy.The data are plotted using Graphpad prism and statistical differencesbetween groups determined.

In the acute setting (3 day), Compound 27 significantly reduced totalprotein, lactate dehydrogenase activity (LDH; tissue injury marker) andtissue inhibitor of metalloproteinase-1 (TIMP-1; pro-fibrotic marker)concentrations in broncheoalveolar lavage fluid (BALF). In the chronicsetting (14 and 28 day) model, Compound 27 maintained mouse body weightand decreased inflammatory cell influx and fibrosis after a singlebleomycin (1.5 units/kg) instillation and decreased pulmonary resistanceand lung fibrosis following repeated bleomycin (3.0-5.0 units/kg/week)instillations.

Compound 37 reduced total protein, lactate and TIMP-1 in the BALF in theacute setting (3-day). Compound 37 decreased inflammatory cell influxand fibrosis after a single bleomycin instillation (3.0 units) in thechronic setting (14-days only).

Example 88 Mouse Carbon Tetrachloride (CCl₄)-Induced Liver FibrosisModel

Female C57BL/6 mice (Harlan, 20-25 g) housed 4/cage are given freeaccess to food and water and allowed to acclimate for at least 7 daysprior to test initiation. After the habituation phase, mice receive CCl₄(0.5-1.0 ml/kg body weight) diluted in corn oil vehicle (100 μL volume)via i.p. injection twice a week for 4-6 weeks. (Higazi, A. A. et al.,Clin Exp Immunol. 2008 April; 152(1):163-73. Epub 2008 Feb. 14). Controlmice receive an equivalent volume of corn oil vehicle only. Testcompound or vehicle is delivered po, ip or sc daily. At the end of thestudy (4-6 weeks after first i.p. injection of CCl₄), mice aresacrificed using inhaled isoflurane and blood is drawn via cardiacpuncture for subsequent analysis of ALT/AST levels. The liver isharvested, and one half of the liver is frozen at −80° C. and the otherhalf is fixed in 10% neutral buffered formalin for histologicalassessment of liver fibrosis using light microscopy (10× magnification).Liver tissue homogenates are analyzed for collagen levels using Sircol(Biocolor Ltd, UK). Fixed Liver tissue is stained using hematoxylin andeosin (H&E) and trichrome and liver fibrosis is determined byquantitative, computer-assisted densitometry of collagen in liver tissuesections using light microscopy. Plasma and liver tissue lysates arealso analyzed for concentrations of inflammatory, pro-fibrotic andtissue injury biomarkers including transforming growth factor β1,hyaluronic acid, tissue inhibitor of metalloproteinase-1, matrixmatelloproteinase-7, connective tissue growth factor and lactatedehydrogenase activity, using commercially available ELISA. Theresulting data are plotted using Graphpad prism and statisticaldifferences between groups determined.

In this experiment, Compound 27 significantly reduced collagendeposition in the liver as compared to the untreated group. Compound 27(30 mg/kg, po, qd) had the same effect on collagen deposition aspirfenidone. Compound 37 significantly reduced collagen deposition inthe liver as compared to untreated control group.

Example 89 Mouse Intravenous LPA-Induced Histamine Release

A mouse intravenous LPA-induced histamine release model is utilized todetermine the in vivo potency of LPA₁ and LPA₃ receptor antagonists.Female CD-1 mice (weighing 25-35 grams) are administered compound (i.p.,s.c. or p.o.) in a volume of 10 ml/kg 30 minutes to 24 hours prior tointravenous LPA challenge (300 μg/mouse in 0.1% FAF BSA) Immediatelyfollowing LPA challenge mice are placed into an enclosed Plexiglaschamber and exposed to an isoflurane for a period of 2 minutes. They areremoved, decapitated and trunk blood collected into tubes containingEDTA. Blood is then centrifuged at 10,000×g for 10 minutes at 4° C.Histamine concentrations in the plasma are determined by EIA. Drugconcentrations in plasma are determined by mass spectrometry. The doseto achieve 50% inhibition of blood histamine release is calculated bynonlinear regression (Graphpad Prism) and plotted as the ED₅₀. Theplasma concentration associated with this dose is plotted as the EC₅₀.

Example 90 Mouse Unilateral Ureteral Obstruction Kidney Fibrosis Model

Female C57BL/6 mice (Harlan, 20-25 g) housed 4/cage will be given freeaccess to food and water and allowed to acclimate for at least 7 daysprior to test initiation. After the habituation phase, mice undergounilateral ureteral obstruction (UUO) surgery or sham to left kidney.Briefly, a longitudinal, upper left incision is performed to expose theleft kidney. The renal artery is located and 6/0 silk thread is passedbetween the artery and the ureter. The thread is looped around theureter and knotted 3 times insuring full ligation of ureter. The kidneyis returned to abdomen, the abdominal muscle is sutured and the skin isstapled closed. Mice are returned to their cages and monitored daily forthe duration of the experiment. Test compound or vehicle is deliveredpo, ip or sc daily. The route and frequency of dosing is based onpreviously determined pharmacokinetic properties. All animals aresacrificed using inhaled isoflurane 4, 8 or 14 days after UUO surgery.Following sacrifice blood is drawn via cardiac puncture, the kidneys areharvested and one half of the kidney is frozen at −80° C. and the otherhalf is fixed in 10% neutral buffered formalin for histologicalassessment of kidney fibrosis using light microscopy (10×magnification). Kidney tissue homogenates are analyzed for collagenlevels using Sircol (Biocolor Ltd, UK). Fixed kidney tissue is alsostained using hematoxylin and eosin (H&E) and trichrome and kidneyfibrosis is determined by quantitative, computer-assisted densitometryof collagen in liver tissue sections using light microscopy. Plasma andkidney tissue lysates are also analyzed for concentrations ofinflammatory, pro-fibrotic and tissue injury biomarkers includingtransforming growth factor β1, hyaluronic acid, tissue inhibitor ofmetalloproteinase-1, matrix matelloproteinase-7, connective tissuegrowth factor and lactate dehydrogenase activity, using commerciallyavailable ELISA. The resulting data are plotted using Graphpad prism andstatistical differences between groups determined

In this experiment, Compound 27 reduced kidney fibrosis by at least 20%as compared to the untreated group. Compound 37 reduced kidney fibrosisby 55% as compared to untreated group.

Example 91 Clinical Trial in Humans with Idiopathic Pulmonary Fibrosis(IPF) Purpose

The purposes of this study is to assess the efficacy of treatment with acompound of Formula (I) compared with placebo in patients withidiopathic pulmonary fibrosis (IPF) and to assess the safety oftreatment with a compound of Formula (I) compared with placebo inpatients with IPF.

The primary outcome variable is the absolute change in percent predictedforced vital capacity (FVC) from baseline to Week 72.

Secondary outcome measures include: composite outcomes of importantIPF-related events; progression-free survival; categorical assessment ofabsolute change in percent predicted FVC from baseline to Week 72;change in Shortness-of-Breath from baseline to Week 72; change inpercent predicted hemoglobin (Hb)-corrected carbon monoxide diffusingcapacity (DLco) of the lungs from baseline to Week 72; change in oxygensaturation during the 6 minute walk test (6MWT) from baseline to Week72; change in high-resolution computed tomography (HRCT) assessment frombaseline to Week 72; change in distance walked in the 6MWT from baselineto Week 72.

Criteria

Patients eligible for this study include those patients that satisfy thefollowing inclusion criteria: diagnosis of IPF; 40 to 80 years of age;FVC≧50% predicted value; DLco≧35% predicted value; either FVC orDLco≦90% predicted value; no improvement in past year; able to walk 150meters in 6 minutes and maintain saturation≧83% while on no more than 6L/min supplemental oxygen.

Patients are excluded from this study if they satisfy any of thefollowing criteria: unable to undergo pulmonary function testing;evidence of significant obstructive lung disease or airwayhyper-responsiveness; in the clinical opinion of the investigator, thepatient is expected to need and be eligible for a lung transplant within72 weeks of randomization; active infection; liver disease; cancer orother medical condition likely to result in death within 2 years;diabetes; pregnancy or lactation; substance abuse; personal or familyhistory of long QT syndrome; other IPF treatment; unable to take studymedication; withdrawal from other IPF trials.

Patients are orally dosed with either placebo or an amount of compoundof Formula (I) (1 mg/day-1000 mg/day). The primary outcome variable willbe the absolute change in percent predicted FVC from Baseline to Week72. Patients will receive blinded study treatment from the time ofrandomization until the last patient randomized has been treated for 72weeks. A Data Monitoring Committee (DMC) will periodically review safetyand efficacy data to ensure patient safety.

After week 72, patients who meet the Progression of Disease (POD)definition, which is a ≧10% absolute decrease in percent predicted FVCor a ≧15% absolute decrease in percent predicted DLco, will be eligibleto receive permitted IPF therapies in addition to their blinded studydrug. Permitted IPF therapies include corticosteroids, azathioprine,cyclophosphamide and N-acetyl-cysteine.

Example 92a Parenteral Pharmaceutical Composition

To prepare a parenteral pharmaceutical composition suitable foradministration by injection (subcutaneous, intravenous, and the like),100 mg of a water-soluble salt of a compound of Formula (I) is dissolvedin sterile water and then mixed with 10 mL of 0.9% sterile saline. Themixture is incorporated into a dosage unit form suitable foradministration by injection

In another embodiment, the following ingredients are mixed to form aninjectable formulation: 1.2 g of a compound of Formulas (I), 2.0 mL ofsodium acetate buffer solution (0.4 M), HCl (1 N) or NaOH (1 M) (q.s. tosuitable pH), water (distilled, sterile) (q.s. to 20 mL). All of theabove ingredients, except water, are combined and stirred and ifnecessary, with slight heating if necessary. A sufficient quantity ofwater is then added.

Example 92b Oral Pharmaceutical Composition

To prepare a pharmaceutical composition for oral delivery, 100 mg of acompound of Formula (I) is mixed with 750 mg of starch. The mixture isincorporated into an oral dosage unit for, such as a hard gelatincapsule, which is suitable for oral administration.

Example 92c Sublingual (Hard Lozenge) Pharmaceutical Composition

To prepare a pharmaceutical composition for buccal delivery, such as ahard lozenge, mix 100 mg of a compound of Formula (I) with 420 mg ofpowdered sugar mixed, with 1.6 mL of light corn syrup, 2.4 mL distilledwater, and 0.42 mL mint extract. The mixture is gently blended andpoured into a mold to form a lozenge suitable for buccal administration.

Example 92d Fast-Disintegrating Sublingual Tablet

A fast-disintegrating sublingual tablet is prepared by mixing 48.5% byweigh of a compound of Formula (I), 44.5% by weight of microcrystallinecellulose (KG-802), 5% by weight of low-substituted hydroxypropylcellulose (50 μm), and 2% by weight of magnesium stearate. Tablets areprepared by direct compression (AAPS PharmSciTech. 2006; 7(2):E41). Thetotal weight of the compressed tablets is maintained at 150 mg. Theformulation is prepared by mixing the amount of compound of Formula (I)with the total quantity of microcrystalline cellulose (MCC) andtwo-thirds of the quantity of low-substituted hydroxypropyl cellulose(L-HPC) by using a three dimensional manual mixer (Inversina®,Bioengineering AG, Switzerland) for 4.5 minutes. All of the magnesiumstearate (MS) and the remaining one-third of the quantity of L-HPC areadded 30 seconds before the end of mixing.

Example 92e Inhalation Pharmaceutical Composition

To prepare a pharmaceutical composition for inhalation delivery, 20 mgof a compound of Formula (I) is mixed with 50 mg of anhydrous citricacid and 100 mL of 0.9% sodium chloride solution. The mixture isincorporated into an inhalation delivery unit, such as a nebulizer,which is suitable for inhalation administration.

In another embodiment, compound of Formula (I) (500 mg) is suspended insterile water (100 mL), Span 85 (1 g) is added followed by addition ofdextrose (5.5 g) and ascorbic acid (10 mg). Benzalkonium chloride (3 mLof a 1:750 aqueous solution) is added and the pH is adjusted to 7 withphosphate buffer. The suspension is packaged in sterile nebulizers.

Example 92f Rectal Gel Pharmaceutical Composition

To prepare a pharmaceutical composition for rectal delivery, 100 mg of acompound of Formula (I) is mixed with 2.5 g of methylcelluose (1500mPa), 100 mg of methylparapen, 5 g of glycerin and 100 mL of purifiedwater. The resulting gel mixture is then incorporated into rectaldelivery units, such as syringes, which are suitable for rectaladministration.

Example 92g Topical Gel Pharmaceutical Composition

To prepare a pharmaceutical topical gel composition, 100 mg of acompound of Formula (I) is mixed with 1.75 g of hydroxypropyl celluose,10 mL of propylene glycol, 10 mL of isopropyl myristate and 100 mL ofpurified alcohol USP. The resulting gel mixture is then incorporatedinto containers, such as tubes, which are suitable for topicaladministration.

Example 92h Ophthalmic Solution

To prepare a pharmaceutical opthalmic solution composition, 100 mg of acompound of Formula (I) is mixed with 0.9 g of NaCl in 100 mL ofpurified water and filtered using a 0.2 micron filter. The resultingisotonic solution is then incorporated into ophthalmic delivery units,such as eye drop containers, which are suitable for ophthalmicadministration.

Example 92i Nasal Spray Solution

To prepare a pharmaceutical nasal spray solution, 10 g of a compound ofFormula (I) is mixed with 30 mL of a 0.05M phosphate buffer solution (pH4.4). The solution is placed in a nasal administrator designed todeliver 100 μl of spray for each application.

The examples and embodiments described herein are for illustrativepurposes only and various modifications or changes suggested to personsskilled in the art are to be included within the spirit and purview ofthis application and scope of the appended claims.

What is claimed is:
 1. A method for treating fibrosis in a mammal inneed of such treatment comprising administering to said mammal, atherapeutically effective amount of a compound of Formula (I), or apharmaceutically acceptable salt thereof:

wherein, R¹ is —CO₂H, —CO₂R^(D), —CN, tetrazolyl, —C(═O)NH₂,—C(═O)NHR¹⁰, —C(═O)NHSO₂R¹⁰ or —C(═O)NHCH₂CH₂SO₃H; R^(D) is H orC₁-C₄alkyl; L¹ is absent or C₁-C₆alkylene; R³ is H, C₁-C₄alkyl,C₃-C₆cycloalkyl, or C₁-C₄fluoroalkyl; R⁷ is H or C₁-C₄alkyl; R⁸ is H,C₁-C₄alkyl, or C₁-C₄fluoroalkyl; R¹⁰ is a C₁-C₆alkyl, C₁-C₆fluoroalkyl,C₃-C₆cycloalkyl, or a substituted or unsubstituted phenyl; each ofR^(A), R^(B), and R^(C) are independently selected from H, F, Cl, Br, I,—CN, —OH, C₁-C₄alkyl, C₁-C₄fluoroalkyl, C₁-C₄fluoroalkoxy, C₁-C₄alkoxy,and C₁-C₄heteroalkyl; m is 0, 1, or 2; n is 0, 1, or 2; and p is 0, 1,or
 2. 2. The method of claim 1, wherein the fibrosis comprises fibrosisof organs or tissues in the mammal.
 3. The method of claim 2, whereinthe fibrosis comprises fibrosis of the heart, kidney, liver, joints,lung, pleural tissue, peritoneal tissue, skin, cornea, retina,musculoskeletal or digestive tract.
 4. The method of claim 1, whereinthe fibrosis is lung fibrosis, kidney fibrosis, liver fibrosis, skinfibrosis, fibrosis of the gut, scleroderma, ocular fibrosis, spinal cordfibrosis, head and neck fibrosis, or myelofibrosis.
 5. The method ofclaim 1, wherein the fibrosis is lung fibrosis, kidney fibrosis, orliver fibrosis.
 6. The method of claim 1, wherein the fibrosis isidiopathic pulmonary fibrosis.
 7. The method of claim 1, wherein thefibrosis is scleroderma.
 8. The method of claim 1, wherein the fibrosisis ocular fibrosis.
 9. The method of claim 1, wherein: R¹ is —CO₂H,—CO₂R^(D) or —C(═O)NHSO₂R¹⁰; R³ is C₁-C₄alkyl; R⁷ is H; R¹⁰ is aC₁-C₆alkyl or a substituted or unsubstituted phenyl; each R^(A) isindependently selected from H, F, Cl, Br, I, —CH₃, —CF₃, —OH, —OCF₃, and—OCH₃; each R^(B) is independently selected from H, F, Cl, Br, I, —CH₃,—CF₃, —OH, —OCF₃, and —OCH₃; each R^(C) is independently selected fromH, F, Cl, Br, I, —CH₃, —CF₃, —OH, —OCF₃, and —OCH₃; m is 0 or 1; and pis 0 or
 1. 10. The method of claim 9, wherein: R¹ is —CO₂H; L¹ isabsent, —CH₂—, —CH₂CH₂—, —CH₂CH₂CH₂—, —CH(CH₃)—, —C(CH₃)₂—,—CH(CH₂CH₃)—, —C(CH₂CH₃)₂—, —CH₂CH(CH₃)—, or —CH₂C(CH₃)₂—; R³ is —CH₃;R⁸ is H, —CH₃ or —CF₃.
 11. The method of claim 10, wherein the compoundof Formula (I) has the following structure:


12. The method of claim 11, wherein: L¹ is absent, —CH₂—, —CH(CH₃)—,—C(CH₃)₂—, —CH(CH₂CH₃)—, or —C(CH₂CH₃)₂—; each R^(C) is independentlyselected from H, F, Cl, —CH₃, —CF₃, —OH, —OCF₃, and —OCH₃; m is 0; n is0, 1, or 2; and p is
 0. 13. The method of claim 10, wherein the compoundof Formula (I) has the following structure:


14. The method of claim 13, wherein: L¹ is absent, —CH₂—, —CH(CH₃)—,—C(CH₃)₂—, —CH(CH₂CH₃)—, or —C(CH₂CH₃)₂—; each R^(C) is independentlyselected from H, F, Cl, —CH₃, —CF₃, —OH, —OCF₃, and —OCH₃; m is 0; n is0, 1, or 2; and p is
 0. 15. The method of claim 9, wherein: R¹ is—C(═O)NHSO₂R¹⁰; R³ is —CH₃ or —CH₂CH₃; R⁸ is H, —CH₃ or —CF₃; and R¹⁰ is—CH₃, or —CH₂CH₃.
 16. The method of claim 1, wherein the compound ofFormula (I) has the following structure:

wherein, R¹ is —CO₂H or —C(═O)NHSO₂R¹⁰; R⁸ is H, or —CH₃; R¹⁰ is aC₁-C₆alkyl or a substituted or unsubstituted phenyl; R^(C) is H, F, Cl,Br, I, —CN, —OH, C₁-C₄alkyl, C₁-C₄fluoroalkyl, C₁-C₄fluoroalkoxy,C₁-C₄alkoxy, or C₁-C₄heteroalkyl.
 17. The method of claim 1, wherein thecompound is:(4′-{3-Methyl-4-[1-(2-trifluoromethyl-phenyl)-ethoxycarbonylamino]-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid (Compound 1);(4′-{3-Methyl-4-[1-(3-trifluoromethyl-phenyl)-ethoxycarbonylamino]-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid (Compound 2);(4′-{4-[1-(2,4-Dichloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid (Compound 3);(4′-{4-[1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid (Compound 4);(4′-{4-[1-(3-Bromo-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid (Compound 5);(4′-{4-[1-(2-Methoxy-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid (Compound 6);(4′-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-6-methoxy-biphenyl-3-yl)-aceticacid (Compound 7);4′-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-carboxylicacid (Compound 8);4′-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-2-carboxylicacid (Compound 9);(4′-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-2-yl)-aceticacid (Compound 10);(4′-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid (Compound 11);(4′-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-3-yl)-aceticacid (Compound 12);3-(4′-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-propionicacid (Compound 13);(4′-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-6-fluoro-biphenyl-3-yl)-aceticacid (Compound 14);(4′-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-4-fluoro-biphenyl-3-yl)-aceticacid (Compound 15);(4′-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid methyl ester (Compound 16);2-(4′-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-propionicacid ethyl ester (Compound 17);2-(4′-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-propionicacid (Compound 18);2-(4′-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-2-methyl-propionicacid (Compound 19);2-(4′-{4-[1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-propionicacid (Compound 20);4-(4′-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-butyricacid (Compound 21);4′-{4-[1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-3-carboxylicacid (Compound 22);(4′-{4-[1-(4-Chloro-2-fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid (Compound 23);(4′-{4-[(R)-1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid (Compound 24);(4′-{4-[(R)-1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-2′-methyl-biphenyl-4-yl)-aceticacid (Compound 25);2-(4′-{4-[1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-2-methyl-propionicacid (Compound 26);(4′-{4-[(R)-1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid (Compound 27);2-(4′-{4-[(R)-1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-2-methyl-propionicacid (Compound 28);2-(4′-{4-[(R)-1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-2-methyl-propionicacid (Compound 29);2-(4′-{4-[(R)-1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-propionicacid (Compound 30);2-(4′-{4-[(R)-1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-propionicacid (Compound 31);(4′-{4-[1-(2,6-Dichloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid (Compound 32);2-(4′-{4-[(R)-1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-2′-methyl-biphenyl-4-yl)-propionicacid (Compound 33);(4′-{4-[(S)-1-(2-Fluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid (Compound 34);(4′-{4-[(S)-1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid (Compound 35);{4′-[4-(2-Chloro-benzyloxycarbonylamino)-3-methyl-isoxazol-5-yl]-biphenyl-4-yl}-aceticacid (Compound 36);{4′-[3-Methyl-4-((R)-1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl]-biphenyl-4-yl}-aceticacid (Compound 37);(4′-{4-[1-(2,3-Difluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid (Compound 38);(4′-{4-[1-(2,4-Difluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid (Compound 39);(4′-{4-[1-(2-Fluoro-4-methoxy-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid (Compound 40);(4′-{4-[1-(2,5-Difluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid (Compound 41);(4′-{4-[1-(2,6-Difluoro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid (Compound 42);{4′-[3-Methyl-4-((R)-1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl]-biphenyl-3-yl}-aceticacid (Compound 43);4′-[3-Methyl-4-((R)-1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl]-biphenyl-4-carboxylicacid (Compound 44);{4′-[3-Methyl-4-((R)-1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl]-biphenyl-2-yl}-aceticacid (Compound 45);{4′-[3-Methyl-4-((R)-1-o-tolyl-ethoxycarbonylamino)-isoxazol-5-yl]-biphenyl-4-yl}-aceticacid (Compound 46);2-(4′-{4-[(R,R)-1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-propionicacid (Compound 47);2-(4′-{4-[(R,S)-1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-propionicacid (Compound 48);(3′-Chloro-4′-{4-[(R)-1-(2-chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid (Compound 49);2-(4′-{4-[(R)-1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-butyricacid (Compound 50);(2′-Chloro-4′-{4-[(R)-1-(2-chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid (Compound 51);(4′-{4-[(R)-1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-2′-fluoro-biphenyl-4-yl)-aceticacid (Compound 52);4′-{4-[(R)-1-(2-Chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-carboxylicacid (Compound 53);(4′-{4-[(R)-1-(3,5-Dibromo-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid (Compound 56);{4′-[3-Methyl-4-((S)-1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl]-biphenyl-4-yl}-aceticacid (Compound 57);(4′-{4-[(R)-1-(3-Hydroxy-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-aceticacid (Compound 58);{4′-[3-Methyl-4-(1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl]-biphenyl-4-yl}-aceticacid (Compound 59);[5-(4′-Cyanomethyl-biphenyl-4-yl)-3-methyl-isoxazol-4-yl]-carbamic acid(R)-1-(2-chloro-phenyl)-ethyl ester (Compound 61);[5-(4′-Cyanomethyl-biphenyl-4-yl)-3-methyl-isoxazol-4-yl]-carbamic acid(R)-1-(2-fluoro-phenyl)-ethyl ester (Compound 62);{3-Methyl-5-[4′-(2H-tetrazol-5-ylmethyl)-biphenyl-4-yl]-isoxazol-4-yl}-carbamicacid (R)-1-(2-fluoro-phenyl)-ethyl ester (Compound 63);{3-Methyl-5-[4′-(2H-tetrazol-5-ylmethyl)-biphenyl-4-yl]-isoxazol-4-yl}-carbamicacid (R)-1-(2-chloro-phenyl)-ethyl ester (Compound 64);[5-(4′-Carbamimidoylmethyl-biphenyl-4-yl)-3-methyl-isoxazol-4-yl]-carbamicacid (R)-1-(2-fluoro-phenyl)-ethyl ester (Compound 65);{5-[4′-(2-Acetylamino-2-imino-ethyl)-biphenyl-4-yl]-3-methyl-isoxazol-4-yl}-carbamicacid (R)-1-(2-fluoro-phenyl)-ethyl ester (Compound 66); or2-(2-{4′-[3-Methyl-4-((R)-1-phenyl-ethoxycarbonylamino)-isoxazol-5-yl]-biphenyl-4-yl}-acetylamino)-ethanesulfonicacid (Compound 67), or a pharmaceutically acceptable salt thereof.